OA11656A - Genes of the 1-deoxy-D-xylulose biosynthetic pathway. - Google Patents

Abstract

The invention relates to the 1-desoxy- D-xylulose- 5-phosphate reductoisomerase gene, the 1-desoxy- D-xylulose- 5-phosphate- synthase gene and the gcpE gene of the 1-desoxy- D-xylulose biosynthetic pathway and to their use for transforming vectors, host organisms and plants and for determining substances that inhibit this biosynthetic pathway.

Description

1 011656

The présent invention relates to DNA sequences which, whenincorporated into the genome of viruses, eukaryotes andprokaryotes, modify isoprenoid biosynthesis and to a geneticengineering process for the production of these transgenicviruses, eukaryotes and prokaryotes. The invention alsorelates to a process for the. identification of substanceshaving herbicidal, antimicrobial, antiparasitic, antiviral,fungicidal, bactericidal action in plants and antimicrobial,antiparasitic, antimycotic, antibacterial and antiviralaction in humans and animais.

The biosynthesis pathway for the formation of isoprenoidsvia the classical acetate/mevalonate pathway and analternative mevalonate-independent biosynthesis pathway, thedeoxy-D-xylulose phosphate pathway is already known (Rohmer, M., Knani, M., Simonin, P., Sutter, B. and Sahm, H. (1993):Biochem. J. 295: 517-524).

It is, . however, not known how and by which pathways it ispossible to bring about a change in the isoprenoidconcentration in viruses, eukaryotes and prokaryotes bymeans of the deoxy-D-xylulose phosphate pathway. Figure 1shows this biosynthesis pathway. DNA sequences are consequently provided which code for 1-deoxy-D-xylulose 5-phosphate synthase (DOXP synthase), 1-deoxy-D-xylulose 5~phosphate reductoisomerase (DOXPreductoisomerase) or the gcpE protein. Ail three genes andenzymes are involved in isoprenoid biosynthesis.

The gcpE protein has a kinase function and catalyses the .phosphorylation of a sugar or a phosphorus sugar or aprecursor of isoprenoid biosynthesis, in particular thephosphorylation of 2-C-methyl-D-erythritol, 2-C-methyl-Derytritol phosphate, in particular 2-C-methyl-D erythritol 2 011656 • 4-phosphate, 2-C-me'thyl-D-erythrose, 2-C-methyl-D-erythrosephosphate, in particular 2-C-methyl-D-erythrose 4-phosphate.•In the precursor of isoprenoid synthesis, the gcpE proteinin particular catalyses the phosphorylation of the following 5 substances: CH2 (OH) -C (CH3) =C (OH) -CH2-O-PO (OH) 2, CH2 (OH)-C (CH3) =C (OH)-CHz-OH, CH2 (OH) -CH (CH3) -CO-CH2-O-PO (OH)2, 10 CH2 (OH) -CH (CH3) -CO-CH2OH, CH2=C (CH3) -CO-CH2-O-PO (OH) 2, CH2=C (CH3) -CO-CH2-OH, CH2=C (CH3)-CH (OH)-CH2-O-PO (OH) 2,, CH2=C (CH3) -CH (OH) -CH2-OH, 15 ,CH2 (0H)-C(=CH2)-C(OH)-CH2-O~PO(OH)2," ' ,CH2 (OH) -C (=CH2) -c (OH) -CH2-OH, CHO-CH (CH3) -CH (OH) -CH2-O-PO- (OH) 2, ' CHO-CH (CH3)-CH (OH)-CH2-OH, CH2(OH) -C(OH) (CH3)-CH=CH-O-PO(OH)2z 20 CH2 (OH) -C (OH) (CH3) -CH=CH-OH, CH(OH)=C (CH3) -CH (OH) -CH2-O-PO(OH)2, CH (OH) =C (CH3) -CH (OH) -CH2-OH, (CH3) 2HC-CO-CH2-O-PO (OH) 2, (CH3) 2hc-co-ch2-o-h, 25 (CH3)2HC-CH(OH) -CH2-O-PO(OH)2, (CH3) 2HC-CH (OH) -CH2-0-H. DOXP synthase catalyses the condensation of pyruvate andglyceraldehyde 3-phosphate to yield 1-deoxy-D-xylulose 5- 30 -phosphate and DOXP reductoisomerase catalyses the conversionof 1-deoxy-D-xylulose 5-phosphate into 2-C-methyl-D-erythritol 4-phosphate (c.f. Fig. 1).

The invention relates to the following DNA sequences: 35 DNA sequences which code for a polypeptide with the a τη) noacid sequence shown in SEQ ID no. 2 or for an analogue ordérivative of the polypeptide according to SEQ.ID no. 2, in 3 011656 which one or more amino acids hâve been deleted, added or.replaced by other amino acids, wherein the enzymatic actionof the polypeptide is retained, and which sequencesoriginate from parasites, wherein sequence variations• occurring within the framework. of natnral strain variabilityare included, DNA sequences which code for a polypeptide with the aminoacid sequence shown in SEQ ID no. 4 or for an analogue ordérivative of the polypeptide according to SEQ ID no. 4, inwhich one or more amino acids hâve been deleted, added orreplaced by other amino acids, wherein the enzymatic actionof the polypeptide is retained, and which sequencesoriginate from parasites, wherein sequence variationsoccurring within the framework of natural strain variabilityare included, and DNA sequences which code for a polypeptide with aminoacid sequence shown in SEQ ID no. 6 or for an analogue ordérivative of the polypeptide according SEQ ID no. 6, inwhich one or more amino acids hâve been deleted, added orreplaced by other amino acids, wherein the catalyticfunction of the polypeptide is retained.

The genes and the gene products thereof (polypeptides) areshown with their primary structure and are assigned asfollows: SEQ ID no. 1: 1-deoxy-D-xylulose 5-phosphatereductoisomerase gene SEQ ID no. 2: 1-deoxy-D-xylulose 5-phosphate reductoisomerase SEQ ID no. 3: 1-deoxy-D-xylulose 5-phosphate synthase SEQ ID no. 4: 1-deoxy-D-xylulose 5-phosphate synthase SEQ ID no. 5: gcpE gene SEQ ID no. 6: gcpE proteins. 4 011656

The DNA sequences a'11 originate from Plasmodium falciparum.

Apart from the DNA sequences stated in the sequence listing,suitable sequences are also those which, as a resuit of thedegeneration of the genetic code, hâve another DNA sequence,but code for the same peptide or for an analogue ordérivative of the polypeptide, in which one or more aminoacids hâve been deleted, added or replaced by other aminoacids.

The sequences according to the invention are suitable forthe expression of genes in viruses, eukaryotes andprokaryotes which are responsible for isoprenoidbiosynthesis in the 1-deoxy-D-xylulose pathway.

According to the invention, eukaryotes or eukaryotic cellsinclude animal cells, plant cells, algae, yeasts, fungi,while prokaryotes or prokaryotic cells include bacteria,archaebacteria and eubacteria.

When a DNA sequence is incorporated into a genome on whichthe above-stated DNA sequence is located, expression of theabove-described genes in viruses, eukaryotes and prokaryotesis enabled. The viruses, eukaryotes and prokaryotestransformed according to the invention are cultivated in amanner known per se and the isoprenoid formed during such•cultivation is isolated and optionally purified. Not ailisoprenoids need to be isolated as in some case theisoprenoids are released directly into the ambient air. 'The invention furthermore relates to a process for theproduction of transgenic viruses, eukaryotes and prokaryotesin order to modify the isoprenoid content, which processcomprises the following steps. a) Production of a DNA sequence with the following sub-sequences i) promoter which is active in viruses,- eukaryotes 5 011656 and prokaryotes and ensures the formation of anRNA in the intended target tissue or target cells, ii) DNA sequence which codes for a polypeptide withthe amino acid sequence shown in SEQ ID no. 2, 4 or 6 or for an analogue or dérivative of thepolypeptide according to SEQ ID no. 2, 4 or 6, iii) 5' and 3' untranslated sequence which enables orenhances expression of the stated genes inviruses, eukaryotes and prokaryotes, b) transfer and incorporation of the DNA sequence into thegenome of viruses, prokarvotic or eukaryotic cells withor without the use of a vector (for example plasmid,viral DNA).

The intact, whole plants may be regenerated from plant cellstransformed in this manner.

The protein-coding sequences with the nucléotide sequencesSEQ ID no. 1, SEQ ID no. 3 and SEQ ID no. 5 may be providedwith a promoter which ensures transcription in certainorgans or cells, which promoter is coupled in senseorientation (3’ end of the promoter to the 5' end of thecoding sequence) to the sequence which codes the protein tobe formed. A termination signal which détermines terminationof mRNA synthesis is attached to the 3' end of the coding'sequence. In order to direct the protein which is to beexpressed to certain subcellular compartments, such aschloroplasts, amyloplasts, mitochondria, vacuoles, cytosolor intercellular spaces, a further sequence which codes fora so-called signal sequence or a transit peptide may beinserted between the promoter and the coding sequence. Insome cases, it is necessary to insert sequences which codefor a signal at the COOH terminus of the protein. Thesequence must be in the same reading frame as the codingsequence of the protein. A large number of cloning vectorsis available in order to préparé for the introduction of theDNA sequences according to the invention into higher plants, 6 011656 which vectors contain a réplication signal for E. coil and amarker which permits sélection of the transformed cells.Depending upon the method by which desired gehes areintroduced into the plant, further DNA sequences may be 5 required. If, for example, the Ti or Ri plasmid is used totransform the plant cells, at least one right border, butfrequently the right border and left border of the Ti and Riplasmid T-DNA must be inserted as a flanking région into the . ’genes to be introduced. The use of T-DNA for transforming10 plant cells has been intensively investigated and .comprehensively described in EP 120516; Hoekama in "TheBinary Plant Vector System", Offset-drukkerij Kanters B.V.Alblasserdam (1985), .chapter V; Fraley et al. Crit.Rev.Plant Sci. 4, 1-46 and An et al. (1985) EMBO J. 4, 277-287. 15 Once the introduced DNA has been incorporated into the genome, it is generally stable and is also retained in thedescendants of the originally transformed cells. It normallycontains a sélection marker, which imparts to thetransformed plant cells résistance to a biocide or an 20 antibiotic, such as kanamycin, G 418, bleomycin, hygromycinor phosphinotricin and others. The particular marker used isthus intended to allow sélection of transformed cells fromcells lacking the inserted DNA. 25 Many techniques are available for introducing DNA into a•plant. These techniques include transformation with theassistance of agrobacteria, for example Agrobacterium tumefaciens, protoplast fusion, microinjection of DNA,electroporation, as well as ballistic methods and virus 30 infection. Whole plants may then be regenerated from thetransformed plant material in a suitable medium which maycontain antibiotics or biocides for sélection purposes. Noparticular requirements are placed upon the plasmids orinjection and electroporation. However, if whole plants are 35 to be regenerated from such transformed cells, a selectablemarker gene must be présent. The transformed cells grow inthe plants in the conventional manner (McCormick et al. 7 011656 (1986), Plant Cell Reports 5, 81-84). The plants may be.cultivated normally and be crossed with plants which hâvethe same transformed genome or other genomes. The résultantindividuals hâve the corresponding phenotypic properties. 5

The présent invention also provides expression vectors whichcontain one or more of the DNA sequences according to the•invention. Such expression vectors are obtained by providingthe DNA sequences according to the invention with suitable 10 functional régulation signais. Such régulation signais are DNA sequences which are responsible for expression, forexample promoters, operators, enhancers, ribosomal bindingsites, and are recognised by the host organism. 15 Further régulation signais, which for example control réplication or recombination of the recombinant DNA in thehost organism, may optionally also be a constituent cart ofthe expression vector. 20 The host organisms transformed with the DNA sequences orexpression vectors according to the invention are alsoprovided by the présent invention.

Suitable host cells and organisms for expressing the enzymes 25 .according to the invention are those which comprise no intrinsic enzymes with the function of DOXP synthase, DOXPreductoisomerase or the gcpE protein. This is the case forarchaebacteria, animais, fungi, slime moulds and someeubacteria. The absence of such intrinsic enzyme activity 30 substantially facilitâtes détection and purification of therecombinant enzymes. As a conséquence, it is also for thefirst time possible straightforwardly to measure, in crudeextracts from the host cells, the activity andin particularthe inhibition of the activity of the recombinant enzymes 35 according to the invention by various Chemicals and pharmaceuticals. 011656

The enzymes according to the invention are advantageouslythen expressed in eukaryotic cells if post-translationalmodification and native folding of the polypeptide chain isto be achieved. Moreover, depending upon the expression 5 System, it is ensured when expressing genomic DNA sequencesthat introns are eliminated by splicing the DNA and theenzymes are produced in the polypeptide sequencescharacteristic to the parasites. Using recombinant DNAtechniques, sequences coding for introns may be eliminated 10 from or inserted for experimental purposes into the DNAsequences to be expressed. • The protein may be isolated from the host cell or theculture supernatant of the host cell using methods known to 15 the person skilled in the art. In vitro réactivation of theenzymes may also be required.

In order to facilitate purification, the enzymes accordingto the invention or sub-sequences of the enzymes may be 20 expressed as fusion proteins with various peptide chains.Oligo-histidine sequences and sequences derived fromglutathione S-transferase, thioredoxin or calmodulin-bindingpeptides are particularly suitable for this purpose. 25 .The enzymes according to the invention or sub-sequences ofthe enzymes may furthermore be expressed as fusion proteinswith such peptide chains known to the person skilled in theart that the recombinant enzymes are transported into theextracellular medium or into certain compartments of the 30 host cells. Both purification and investigation of thebiological activity of the. enzymes may consequently befacilitated.

When expressing the enzymes according to the invention, it 35 may prove convenient to modify individual codons. Purposefulreplacement of bases in the coding région may here also beadvisable if the codons used in the parasites differ from 011656 the codon use in the heterologous expression System, in order to ensure optimal synthesis of the protein. •The enzymes according to the invention may furthermore be 5 obtained under standardised conditions by in vitro translation by methods known to the person skilled in the'art. Systems suitable for this purpose are rabbitréticulocyte and wheat germ extracts and bacterial lysâtes.In vitro transcribed mRNA may also be translated into 10 . Xenopus oocytes. .Oligo- and polypeptides, the sequences of which are derivedfrom the peptide sequence of the enzymes according to theinvention, may be obtained by Chemical synthesis. Given 15 appropriate sélection of the sequences, such peptides hâveproperties which are characteristic of the enzymes accordingto the invention. Such peptides may be produced in largequantifies and are particularly suitable for investigatingthe kinetics of enzyme activity, régulation of enzyme 20 activity, the threedimensional structure of the enzymes,inhibition of enzyme activity by various Chemicals andpharmaceuticals and the binding geometry and bindingaffinity of various ligands. 25 · DNA with the nucléotides from sequences SEQ ID no. 1, 3 and5 are preferably used for the recombinant production of the •enzymes according to the invention.

The invention accordingly moreover relates to a process for

30 screening for compounds which inhibit the deoxy-D-xylulosephosphate metabolic pathway. According to this process, ahost organism, which contains a recombinant expressionvector, wherein the vector comprises at least a portion ofthe oligonucleotide sequence according to SEQ ID no. 1, SEQ 35 ID no. 3 or SEQ ID no. 5 or variants or homologues thereof,is provided, as is a compound which is suspected to hâveantimicrobial, antiparasitic, antibacterial, antiviral and - 10 011656 antimycotic action in humans and animais or an 'antimicrobial, antiviral, bactericidal, herbicidal orfungicidal activity in plants. The host organism is thenbrought into contact 'with the compound and the activity of.the compound determined.

The présent invention also provides methods for determiningthe enzymatic activity of the gcpE protein. Said activitymay be determined using known methods. Détermination isperformed by detecting the phosphorylation of a sugar or of•a phosphorus sugar or of a precursor of isoprenoidbiosynthesis, in particular the phosphorylation of 2-C-methyl-D-erythritol, 2-C-methyl-D-erytritol phosphate, in•particular 2-C-methyl-D-erythritol 4-phosphate, 2-C-methyl-D-erythrose, 2-C-methyl-D-erythrose phosphate, in particular 2-C-methyl-D-erythrose 4-phosphate. The présent inventionalso provides the use of this measurement method foridentifying substances which inhibit the activity of theparticular enzymes.

The enzymatic activity of DOXP synthase and DOXPreductoisomerase may be detected in a single step bydetermining the conversion of glyceraldehyde 3-phosphateinto 2-C-methylerythritol 4-phosphate. 'Détermination of the activities of DOXP synthase and DOXPreductoisomerase proceeds analogously. Fluorimetric methodsdescribed by Querol et al, are also suitable for determining‘DOXP synthase activity (Querol at al., abstracts, 4thEuropean Symposium on Plant Isoprenoids, Barcelona, 21-23.April 1999).

Claims (7)

11 C116 56 Claims 10 DNA sequences which code for a polypeptide with theamino acid sequence shown in SEQ ID no. 2 or for ananalogue or dérivative of the polypeptide according toSEQ ID no. 2, in which one or more amino acids hâvebeen deleted, added or replaced by other amino acids,wherein the enzymatic action of the polypeptide isretained, and which sequences originate fromparasites, wherein sequence variations occurringwithin the framework of naturel strain variability areincluded. • 2. 15 20 25 3. 30 4 35 DNA sequences which code for a polypeptide with theamino acid sequence shown in SEQ ID no. 4 or for ananalogue or dérivative of the polypeptide according toSEQ ID no. 4, in which one or more amino acids hâvebeen deleted, added or replaced by other amino acids,wherein the enzymatic action of the polypeptide isretained, and which sequences originate fromparasites, wherein sequence variations occurringwithin the framework of natural strain variability areincluded. DNA sequences which code for a polypeptide with theamino acid sequence shown in SEQ ID no. 6 or for ananalogue or dérivative of the polypeptide according toSEQ ID no. 6, in which one or more amino acids hâvebeen deleted, added or replaced by other amino acidswherein the catalytic function of the polypeptide isretained. DNA sequence according to one of claims 1 to 3,characterised in that it also comprises functionalrégulation signais, in particular promoters,operators, enhancers, ribosomal binding sites. 12 011656 .5. DNA sequence with the following sub-sequences i) promoter which is active in viruses, eukaryotes and prokaryotes and ensures the formation of anRNA in the intended target tissue or target cells, 5 ii) DNA sequences according to one of daims 1 to 3, iii)3' untranslated sequence which, in viruses, eukaryotes and prokaryotes, results in theaddition of poly(A) residues onto the 3' end ofthe RNA. 10

6. Process for the production of transgenic viruses,eukaryotes and prokaryotes for modifying theisoprenoid content, characterised in that a DNAsequence according to claim 4 or 5 is transferred and 15 incorporated into the genome of viruses, eukaryotic and prokaryotic cells with or without use of a vector.

7. Transgenic Systems, in particular plants and plantcells which contain one or more DNA sequences 20 according to daims 1 to 5 as '"foreign" or "additional" DNA, which sequences are expressed.

8. Expression vector containing one or more DNA sequencesaccording to daims 1 to 5. 25

9. Protein which is involved in the 1-deoxy-D-xylulose 5-phosphate metabolic pathway and a) is coded by DNAsequences SEQ ID no. 1, 3 or 5 or b) is coded by DNAsequences which hybridise with DNA sequences SEQ ID 30 no.1, 3, 5 or fragments of these DNA sequences in the DNA région which codes for the mature protein. .10. Protein according to claim 9, obtainable from the culture supernatants of parasites or from the 35 disrupted parasites and purification by chromatographie and electrophoretic methods. 13 011656 Protein according to one of daims 9 and 10, characterised in that it a) is the product of viral,prokaryotic or eukaryotic expression of exogenous DNA,b) is coded by sequences SEQ ID no. 1, 3 or 5 or iscoded by DNA sequences which hybridise with DNAsequences SEQ ID no. 1, 3, 5 or fragments of these DNAsequences in the DNA région which codes for the matureprotein, or c) is coded by DNA sequences which wouldhybridise without degeneration of the genetic codewith the sequences defined in b) and which code for apolypeptide with a corresponding amino acid sequence. Protein according to one of the preceding daims,characterised in that it comprises the amino acidsequences SEQ ID no. 2, 4 or 6. Process for determining the enzymatic activity of thegcpE protein, characterised in that phosphorylation ofa sugar or of a phosphorus sugar or of a precursor ofisoprenoid biosynthesis, in particular the phosphorylation of 2-C-methyl-D-erythritol, 2-C-methyl-D-erytritol phosphate, in particular 2-C-methyl-D-erythritol 4-phosphate, 2-C-methyl-D-erythrose, 2-C-methyl-D-erythrose phosphate, inparticular 2-C-methyl-D-erythrose 4-phosphate, and ofphosphate and alcohol precursors, is detected. Process according to daim 13, characterised in thatphosphorylation of the following phosphates oralcohols is detected: CH2 (OH) -C (CH3) =C (OH) -CH2-O-PO (OH) 2, CH2 (OH) -C (CH3) =C (OH) -CH2-OH, CH2 (OH) -CH (CH3) -CO-CH2-O-PO (OH) 2, CHZ (OH)-CH (CH3)-CO-CH2OH, CH2=C (CH3)-CO-CH2-O-PO (OH) 2, CH2=C (CH3)-CO-CHz-OH, CH2=C (CH3) -CH (OH) -CH2-O-PO (OH) 2, 14 011656 CHZ=C (CH3) -CH (.OH) -CHz-OH, · CH2 (OH) -C (=CH2) -C (OH) -CH2-O-PO(OH) 2,CH2 (OH) -C (=CH2) -C (OH) -CH2-OH,CH0-CH(CH3) -CH (OH) -CH2-O-PO-(OH)2, 5 CHO-CH(CH3)-CH(OH)-CH2-OH, CH2 (OH) -C (OH) (CH3) -CH=CH-O-PO (OH) 2,CH2 (OH)-C (OH) (CH3)-CH=CH-OH, CH (OH) =C (CH3) -CH (OH) -CH2-O-PQ (OH) 2,CH (OH) =C (CH3) -CH (OH) -CH2-OH, 10 (CH3)2HC-CO-CH2-O-PO(OH)2, (CH3) 2hc-co-ch2-o-h, (CH3) 2HC-CH (OH)-CH2-O-PO (OH) 2, (CH3) 2HC-CH (OH) -CH2-0-H. 15 15 20 16 25 30 35 Process for the combined détermination of theenzymatic activity of DOXP synthase and of DOXPreductase, characterised in that the conversion ofglyceraldehyde 3-phosphate into 2-C-methyl erythritol4-phosphate is detected. Process for screening a compound for the treatment ofinfectious processes in humans and animais, whereinthe process comprises: a) provision of a host cell which contains arecombinant expression vector, wherein the vectorcomprises at least a portion of the oligonucleotide sequence according to SEQ ID no.1, SEQ ID no. 3 or SEQ ID no. 5 or variants oranalogues thereof, and moreover of a compoundsuspected to hâve antimycotic, antibiotic,antiparasitic or antiviral action in humans andanimais, b) bringing the host cell into contact with thecompound and c) determining the antimicrobial, antimycotic,antibiotic, antiparasitic or antiviral action ofthe compound. 15 011656

17. Process for screening for compounds for treatingplants, wherein the process comprises: a) provision of a host cell which contains a 5 recombinant expression vector, wherein the vector comprises' at least a portion of theoligonucleotide sequence according to SEQ ID no.1, SEQ ID no. 3 or SEQ ID no. 5 or variants oranalogues thereof, and moreover of a compound 10 suspected to hâve antimicrobial, antiviral, antiparasitic, bactericidal, fungicidal orherbicidal action in plants, b) bringing the host cell into contact with thecompound and 15 c) determining the antimicrobial, antiviral, antiparasitic, bactericidal, fungicidal orherbicidal action of the compound.

18. Use of DNA according to one of daims 1 to 5 or ofproteins according to one of daims 9 to 12 or oftransgenic Systems according to daim 7 for theprévention or treatment of diseases in humans andanimais. 20 -1 - 011656 SEQUENCE LISTING <110> Jomaa, Hassan <120> Genes of the 1-deoxy-D-xylulose biosynthesis pathway <130> 15696 <140> PCT/EP99 <141> 1999-09-22 <150> DE19923567.8 <151> 1999-05-22 <150> DE19343279.8 <151> 1998-09-22 <160> 6 <170> Patentln Ver. 2.1 <210> 1 <211> 146“ <212> DNA <213> Plasmodium falciparum <220> <221> CCS <222> ( i> . . (14 67) <220> <221> gens <222> (1) . . (14 67) <220> <221> mP.NA <222> (1)..(1467) <400> 1 atg Met 1 aag Lys 3câ Lys tat Tyr att Ile 5 tat Tyr ata Ile tat Tyr ttt Phe ttc Phe 10 ttô Phe atc Ile aca Thr ata Ile act Thr 15 att Ile 48 aat gat tta gta ata aat aat aca tca aaa tgt gtt tcc att gaa aga 96 Asn Asp Leu Val 20 Ile Asn Asn Thr Ser 25 Lys Cys Val Ser Ile 30 Glu Arg aga aaa aat aac gca tat ata aat tat ggt ata gga tat aat gga cca 144 Arg Lys Asn 5 K· Asn Al a Tyr Ile Asn 40 Tyr Gly Ile Gly Tyr 45 Asn Gly Pro gat aat O.Ô.Ô. ata aca aag agt aga aga tgt aaa aga ata aag tta tgc 192 Asp Asn 50 Lys Ile Thr Lys Ser 55 Arg Arg Cys Lys Arg 60 Ile Lys Leu Cys -2- 011656 aaa Lys 65 aag Lys gat Asp tta Leu ata Ile qat Asp 70 att Ile ggt Gly gca Ala ata Ile aag Lys 75 aaa Lys cca Pro att Ile aat Asn gta Val 80 240 gca att ttt gga agt act ggt agt ata ggt acg aat gct tta aat ata 288 Ala Ile Phe Gly Ser 85 Thr Gly Ser Ile Gly 90 Thr Asn Ala Leu Asn 95 Ile ata agg gag tgt aat aaa att gaa aàt gtt ttt aat gtt aaa gca ttg 336 Ile Arg Glu Cys 100 Asn Lys Ile Glu Asn 105 Val Phe Asn Val Lys 110 Ala Leu tat gtg aat aag agt gtg aat qaa tta tat gaa caa gct aga gaa ttt 384 Tyr Val Asn 115 Lys Ser Val Asn Glu 120 Leu Tyr Glu Gin Ala 125 Arg Glu Phe tta cca gaa tat ttg tgt ata cat gat aaa agt gta tat gaa gaa tta 432 Leu Pro 130 Glu Tyr Leu Cys Ile 135 His Asp Lys Ser Val 140 Tyr Glu Glu Leu aaa gaa ctg gta aaa aat ata aaa gat tat aaa cct ata ata ttg tgt 480 Lys 145 Glu Leu Val Lys Asn 150 Ile Lys Asp Tyr Lys 155 Pro Ile Ile Leu Cys 160 ggt gat gaa ggg atg aaa gaa ata tgt agt agt aat agt ata gat aaa 528 Gly Asp Glu Gly Met 165 Lys Glu Ile Cys Ser 170 Ser Asn Ser Ile Asp 175 Lys ata gtt cfC ~ ggt att gat tct ttt caa gga tta tat tct act atg tat 576 Ile Val Ile Gly 180 Ile Asp Ser Phe Gin 185 Gly Leu Tyr Ser Thr 190 Met Tyr gca att at 5 aat aat aaa ata gtt gcg tta gct aat aaa gaa tcc att 624 Ala Ile Mer 195 Asn Asn Lys Ile Val 200 Ala Lèu Ala Asn Lys 205 Glu Ser Ile gtc tct gcr ggt ttc ttt tta aag aaa tta tta aat att cat aaa aat 672 Val Ser 210 àla Gly Phe Phe Leu 215 Lys Lys Leu Leu Asn 220 Ile His Lys Asn gca aag ata ata cct gtt gat tca gaa cat agt gct ata ttt caa tgt 720 Al a225 Lys Ile Ile Pro Val 230 Asp Ser Glu His Ser 235 Ala Ile Phe Gin Cys 240 tta gat aat aat aag gta tta aaa aca aaa tgt tta caa gac aat ttt 768 Leu Asp Asn Asn Lys 245 Val Leu Lys Thr Lys 250 Cys Leu Gin Asp Asn 255 Phe tct aaa att aac aat ata aat aaa ata ttt tta tgt tca tct gga ggt 816 Ser Lys Ile Asn 260 Asn Ile Asn Lys Ile 265 Phe Leu Cys Ser Ser 270 Gly Gly cca ttt caa aat tta act atg gac gaa tta aaa aat gta aca tca gaa 864 Pro Phe Gin27 5 Asn Leu Thr Met Asp 280 Glu Leu Lys Asn Val 285 Thr Ser Glu 011656 aat Asn gct Ala 290 tta Leu aag Lys cat His cct Pro aaa Lys 295 tgg Trp aaa Lys atg Met ggt Gly aag Lys 300 aaa Lys ata Ile act Thr ata Ile 912 gat tct gca act atg atg aat aaa ggt tta gag gtt ata gaa acc cat 960 Asp Ser Ala Thr Met Met Asn Lys. Gly Leu Glu Val Ile Glu Thr His 305 310 315 320 ttt tta ttt gat gta gat tat aat gat ata gaa gtt ata gta cat aaa 1008 Phe Leu Phe Asp Val Asp Tyr Asn Asp Ile Glu Val Ile Val His Lys 325 330 335 gaa tgc att ata cat tct tgt gtt gaa ttt ata gac aaa tca gta ata 1056 Glu Cys Ile Ile His Ser Cys Val Glu Phe Ile Asp Lys Ser Val Ile 340 345 350 agt caa atg tat tat cca gat atg caa ata ccc ata tta tat tct tta 1104 Ser Gin Met Tyr Tyr Pro Asp Met Gin Ile Pro Ile Leu Tyr Ser Leu 355 360 365 aca tgg cct gat aga ata aaa aca aat tta aaa cct tta gat ttg gct 1152 Thr Trp Pro Asp Arg Ile Lys Thr Asn Leu Lys Pro Leu Asp Leu Ala 370 375 380 cag gtt act c.tt aca ttt cat aaa cct tct tta gaa cat ttc ccg 1200 Gin Val Ser Thr Leu Thr Phe His Lys Pro Ser Leu Glu His Phe Pro 385 390 395 400 tgt att 3 3 3 tta gct tat caa gca ggt ata aaa gga aac ttt tat cca 1248 Cys Ile L S Leu Ala Tyr Gin Ala Gly Ile Lys Gly Asn Phe Tyr Pro 405 410 415 act gta CC 3 aat gcg tca aat gaa ata gct aac aac tta ttt ttg aat 1296 Thr Val Leu Asn Ala Ser Asn Glu Ile Ala Asn Asn Leu Phe Leu Asn 420 425 430 aat aaa 3 L U aaa tat ttt gat att tcc tct ata ata tcg caa gtt ctt 1344 Asn Lys Ile Lys Tyr Phe Asp Ile Ser Ser Ile Ile Ser Gin Val Leu 4 35 440 445 gaa tct ttc aat tct caa aag gtt tcg gaa aat agt gaa gat tta atg 1392 Glu Ser Phe Asn Ser Gin Lys Val Ser Glu Asn Ser Glu Asp Leu Met 450 455 460 aag caa att cta caa ata cat tct tgg gcc aaa gat aaa gct acc gat 1440 Lys Gin Ile’ Leu Gin Ile His Ser Trp Ala Lys Asp Lys Ala Thr Asp 465 470 475 480 ata tac aac aaa cat aat tct tca tag 1467 Ile Tyr Asn Lys His Asn Ser Ser <210> 2 <211> 488 <212> PRT <213> Plasmodium falciparum 485 -4- 011656 <400> 2 Phe 10 Phe Ile Thr Ile Thr 15 Ile Met 1 Lys Lys Tyr Ile 5 Tyr Ile Tyr Phe Asn Asp Leu Val Ile Asn Asn Thr Ser Lys Cys Val Ser Ile Glu Arg 20 25 30 Arg Lys Asn Asn Ala Tyr Ile Asn Tyr Gly Ile Gly Tyr Asn Gly Pro 35 40 45 Asp Asn Lys Ile Thr Lys Ser Arg Arg Cys Lys Arg Ile Lys Leu Cys 50 55 60 Lys Lys Asp Leu Ile Asp Ile Gly Ala Ile Lys Lys Pro Ile Asn Val 65 70 75 80 Al a Ile Phe Gly Ser Thr Gly Ser Ile Gly Thr Asn Ala Leu Asn Ile 85 90 95 Ile Arg Glu Cys Asn Lys Ile Glu Asn Val Phe Asn Val Lys Ala Leu 100 105 110 Tyr Val Asn Lys Ser Val Asn Glu Leu Tyr Glu Gin Ala Arg Glu Phe 115 120 125 Leu Pro Glu Tyr Leu Cys Ile His Asp Lys Ser Val Tyr Glu Glu Leu 130 135 140 Lys Glu Leu Val Lys Asn Ile Lys Asp Tyr Lys Pro Ile Ile Leu Cys 145 150 155 160 Gly Asp Glu Gly Met Lys Glu Ile Cys Ser Ser Asn Ser Ile Asp Lys 165 170 175 Ile Val Ile Gly Ile Asp Ser Phe Gin Gly Leu Tyr Ser Thr Met Tyr 180 185 190 Al a Ile Met Asn Asn Lys Ile Val Ala Leu Ala Asn Lys Glu Ser Ile 195 200 205 Val Ser Ala Gly Phe Phe Leu Lys Lys Leu Leu Asn Ile His Lys Asn 210 215 220 Al a Lys Ile Ile Pro Val Asp Ser Glu His Ser Ala Ile Phe Gin Cys 225 230 235 240 Leu Asp Asn Asn Lys Val Leu Lys Thr Lys Cys Leu Gin Asp Asn Phe 245 250 255 Ser Lys Ile Asn Asn Ile Asn Lys Ile Phe Leu Cys Ser Ser Gly Gly 260 265 270 Pro Phe Gin Asn Leu Thr Met Asp Glu Leu Lys Asn Val Thr Ser Glu 275 280 285 Asn Al a Leu Lys His Pro Lys Trp Lys Met Gly Lys Lys Ile Thr Ile 290 295 300 011656 Asp 305 Ser Al a Thr Met Met 310 Asn Lys Gly Leu Glu315 Val Ile Glu Thr His 320 Phe Leu Phe Asp Val Asp Tyr Asn Asp Ile Glu Val Ile Val His Lys 325 330 335 Glu Cys Ile Ile His Ser Cys Val Glu Phe Ile Asp Lys Sér Val Ile 340 345 350 Ser Gin Mer Tyr Tyr Pro Asp Met Gin Ile Pro Ile Leu Tyr Ser Leu 355 360 365 Thr Trp Pro Asp Arg Ile Lys Thr Asn Leu Lys Pro Leu Asp Leu Ala 370 375 380 Gin Val Ser Thr Leu Thr Phe His Lys Pro Ser Leu Glu His Phe Pro 385 390 395 400 Cys Ile Lys Leu Ala Tyr Gin Ala Gly Ile Lys Gly Asn Phe Tyr Pro 405 410 415 Thr Val Leu Asn Ala Ser Asn Glu Ile Ala Asn Asn Leu Phe Leu Asn 420 425 430 Asn Lys Ile Lys Tyr Phe Asp Ile Ser Ser Ile Ile Ser Gin Val Leu 4 35 440 445 Glu Ser Phe Asn Ser Gin Lys Val Ser Glu Asn Ser Glu Asp Leu Met 450 455 460 Lys Gin Ile Leu Gin Ile His Ser Trp Ala Lys Asp Lys Ala Thr Asp 465 470 475 480 Ile Tyr Asn Lys His Asn Ser Ser 485 <210> 3 <211> 3872 <212> DNA <213> Plas-.odium falciparum <220> <221> CDS <222> (126)..(3740) <220> <221> gene <222> (1) . .(3870) <220> <221> mRNA <222> (1)..(3870) <400> 3 ggtaatatac atataatata tatataatat attcttacgt atgtatcatt tatgaatcat 60 -6- 011656 aataatattc taaatttacc ttccgttttt gctcgatctt ctcattttcg tttcagcttt 120 tatca atg att ttt aat tat gtg ttt ttt aag aac ttt gta cca gtt gtt 170Met Ile Phe Asn Tyr Val Phe Phe Lys Asn Phe Val Pro Val Val 15 10 15 cta Leu tac Tyr att Ile etc Leu ctt Leu 20 ata Ile ata Ile tat Tyr att Ile aac Asn 25 tta Leu aat Asn ggc Gly atg Met aat Asn 30 aat Asn 218 aaa aat caa ata aaa aca gaa aaa att tat ata aag aaa ttg aat agg 266 Lys Asn Gin Ile 35 Lys Thr Glu Lys Ile 40 Tyr Ile Lys Lys Leu 45 Asn Arg ttg tca agg aaa aat teg tta tgt agt tet aaa aat aaa ata gea tgc 314 Leu Ser Ara 50 Lys Asn Ser Leu Cys 55 Ser Ser Lys Asn Lys 60 Ile Al a Cys ttg ttc gat ata gga aat gat gat aat aga aat aeg aca tat ggc tat 362 Leu Phe 65 Asp Ile Gly Asn Asp 70 Asp Asn Arg Asn Thr 75 Thr Tyr Gly Tyr aat gta âüL gtt aaa aat gat gat att aat tcc tta cta aaa aat aat 410 Asn 80 Val Asn Val Lys Asn 85 Asp Asp Ile Asn Ser 90 Leu Leu Lys Asn Asn 95 tat agt aaa ttg tac atg gat aag agg aaa aat att aat aat gta 458 Tyr Ser Asn Lys Leu 100 Tyr Met Asp Lys Arg 105 Lys Asn Ile Asn Asn 110 Val att agt 5.3“ aat aaa ata tet ggg tcc att tca aat att tgt agt aga 506 Ile Ser îr.r Asn 115 Lys Ile Ser Gly Ser 120 Ile Ser Asn Ile Cys 125 Ser Arg aat caa âc = gaa aat gaa caa aaa aga aat aaa caa aga tgt tta act 554 Asn Gin Lys 130 Glu Asn Glu Gin Lys 135 Arg Asn Lys Gin Arg 140 Cys Leu Thr caa tgt cac act tat aat atg tca cat gaa cag gac aaa cta gct aat 602 Gin Cys 145 Hrs Thr Tyr Asn Met 150 Ser His Glu Gin Asp 155 Lys Leu Ala Asn gat aat agg aat aat aaa aag aat ttt aat tta tta ttt ata aat 650 Asp 160 Asn Asn Arg Asn Asn 165 Lys Lys Asn Phe Asn 170 Leu Leu Phe Ile Asn 175 tat ttt aat ttg aaa ega atg aaa aat tet ctt cta aat aaa gac aat 698 Tyr Phe Asn Leu Lys 180 Arg Met Lys Asn Ser 185 Leu Leu Asn Lys Asp 190 Asn ttc ttt tac tgt aaa gaa aaa aaa ttg tca ttt ctg cat aag gcc tat 746 Phe Phe Tyr Cys 195 Lys Glu Lys Lys Leu 200 Ser Phe Leu His Lys 205 Ala Tyr aaa aaa aaa aat tgc act ttt caa aat tat agt tta aaa aga aaa tet 794 Lys Lys Lys 210 Asn Cys Thr Phe Gin 215 Asn Tyr Ser Leu Lys 220 Arg Lys Ser 7- 011656 aat Asn cgt Arg 225 gat Asp tca Ser cat His aaa Lys ttg Leu 230 ttt Phe tct Ser gga Gly gaa Glu ttt Phe 235 gac Asp gat Asp tat Tyr aca Thr 842 aat aat aat gct tta tat gaa tcc gaa aaa aaa gaa tac att aca cta 890 Asn Asn Asn Ala Leu Tyr Glu Ser Glu Lys Lys Glu Tyr Ile Thr Leu 240 245 250 255 aat aat aat aat aaa aat aat aat aat aaa aat aat gat aat aaa aat 938 Asn Asn Asn. Asn Lys Asn Asn Asn Asn Lys Asn Asn Asp Asn Lys Asn 260 265 270 aat gat aat aat gat tat aat aat aat aat agt tgt aat aat tta gga 986 Asn Asp Asn Asn Asp Tyr Asn Asn Asn Asn Ser Cys Asn Asn Leu Gly 275 280 285 gag aga tcc aat cat tat gat aat tat ggt gga gat aat aat aat cca 1034 Glu Arg Ser Asn His Tyr Asp Asn Tyr Gly Gly Asp Asn Asn Asn Pro 290 295 300 tgt aat aSTL aat aat gac aaa tat gat ata gga aaa tat ttc aaa cag , 1082 Cys Asn Asn Asn Asn Asp Lys Tyr Asp Ile Gly Lys Tyr Phe Lys Gin 305 310 315 att aat aoc ttt att aat att gat gaa tat aaa act ata tat ggt gat 1130 Ile Asn Thr Phe Ile Asn Ile Asp Glu Tyr Lys Thr Ile Tyr Gly Asp 320 325 330 335 gaa ata tat aaa gaa ata tat gaa cta tat gta gaa aga aat att cct 1178 Glu Ile Lys Glu Ile Tyr Glu Leu Tyr Val Glu Arg Asn Ile Pro 340 345 350 gaa tat *57 gaa cga aaa tat ttt tca gaa gat att aaa aag agt gtc 1226 Glu Tyr T ·. .· ** Glu Arg Lys Tyr Phe Ser Glu Asp Ile Lys Lys Ser Val 355 360 365 cta ttt ata gat aaa tat aat gat gtc gaa ttt gaa aaa gct ata 1274 Leu Phe HS C Ile Asp Lys Tyr Asn Asp Val Glu Phe Glu Lys Ala Ile 3“0 375 380 aaa gaa ÇâS ttt ata aat aat gga gtt tat att aat aat ata gat aat 1322 Lys Glu Gi;: Phe Ile Asn Asn Gly Val Tyr Ile Asn Asn Ile Asp Asn 385 390 395 aca tat tat aaa aaa gaa aat att tta ata atg aaa aag ata tta cat 1370 Thr Tyr Tyr Lys Lys Glu Asn Ile Leu Ile Met Lys Lys Ile Leu His 400 405 410 415 tat ttc CCg tta tta aaa tta att aat aat cca tca gat tta aaa aag 1418 Tyr Phe Prc Leu Leu Lys Leu Ile Asn Asn Pro Ser Asp Leu Lys Lys 420 425 430 tta aaa aaa caa tat tta cct tta tta gca cat gaa tta aaa ata ttt 1466 Leu Lys Lys Gin Tyr Leu Pro Leu Leu Ala His Glu Leu Lys Ile Phe 435 440 445 8 - 011656 tta Leu ttt Phe ttt Phe l450 att Ile gta Val aat Asn ata Ile aca Thr 455 gga Gly ggt Gly cat His ttt Phe tcc Ser 460 tct Ser gtt Val tta Leu 1514 agc tct tta gaa att caa tta tta tta ttg tat att ttt aat caa cca 1562 Ser Ser Leu Glu Ile Gin Leu Leu Leu Leu Tyr Ile Phe Asn Gin Pro 465 470 475 tat gat aat gtt ata tat gat ata gga cat caa gca tat gta cat aag 1610 Tyr Asp Asn Val. Ile Tyr Asp Ile Gly His Gin Ala Tyr Val His Lys 480 485 490 495 ata ttg ace gga aga aaa cta tta ttt cta tca tta aga aat aaa aaa 1658 Ile Leu Thr Gly Arg Lys Leu Leu Phe Leu Ser Leu Arg Asn Lys Lys 500 505 510 ggt att a e~ gga ttc cta aat att ttt gaa agt att tat gat aaa ttt 1706 Gly Ile Ser Gly Phe Leu Asn Ile Phe Glu Ser Ile Tyr Asp Lys Phe 515 520 525 ggg gct ace cac agt tcc act tca tta agt gct ata caa gga tat tat 1754 Gly Ala Gly His Ser Ser Thr Ser Leu Ser Ala Ile Gin Gly Tyr Tyr 5 Ξ 5 535 540 gaa. gcc aaa tgg caa gtg aag aat aaa gaa aaa tat gga aat gga gat 1802 Glu Ala Trp Gin Val Lys Asn Lys Glu Lys Tyr Gly Asn Gly Asp 545 550 555 ata gaa £ Σ £. agt gat aac gca aat gtc acg aat aat gaa agg ata ttt 1850 Ile Glu Ile Ser Asp Asn Ala Asn Val Thr Asn Asn Glu Arg Ile Phe 560 565 570 575 caa aaa gça ata cac aat gat aat aat att aac aat aat att aat aat 1898 Gin Lys Gly Ile His Asn Asp Asn Asn Ile Asn Asn Asn Ile Asn Asn 580 585 590 aat aat tat atc aat cct tca gat gtg gta gga aga gaa aat acg aat 1946 Asn Asn T - · >- Ile Asn Pro Ser Asp Val Val Gly Arg Glu Asn Thr Asn 595 600 605 gta cca âïZ gta cga aat gat aac cat aac gtg gat aaa gta cac att 1994 Val Pro as n Val Arg Asn Asp Asn His Asn Val Asp Lys Val His Ile 610 615 620 gct att £ ~ 5. gga gat ggt ggt tta aca ggt gga atg gca tta gaa gcg 2042 Al a Ile Ile Gly Asp Gly Gly Leu Thr Gly Gly Met Ala Leu Glu Ala 625 630 635 tta aat t £ ~ att tca ttc ttg aat tct aaa att tta att att tat aat 2090 Leu Asn Tyr Ile Ser Phe Leu Asn Ser Lys Ile Leu Ile Ile Tyr Asn 640 645 650 655 gat aac gça caa gtt tct tta cca aca aat gcc gta agt ata tca ggt 2138 Asp Asn Gly Gin Val Ser Leu Pro Thr Asn Ala Val Ser Ile Ser Gly 660 665 670 9- 011656 aat Asn aga Arg cct Pro ata Ile 675 ggt Gly tct Ser ata Ile tca Ser gat Asp 680 cat His tta Leu cat His tat Tyr ttt Phe 685 gtt Val tctSer ' 2186 aat ata gaa gea aat gct ggt gat aat aaa tta tcg aaa aat gea aaa 2234 Asn Ile Glu Al a Asn Ala Gly Asp Asn Lys Leu Ser Lys Asn Ala Lys 690 695 700 gag aat aac att ttt gaa aat ttg aat tat gat tat att ggt gtt gtg 2282 Glu Asn Asn Ile Phe Glu Asn Leu Asn Tyr Asp Tyr Ile Gly Val Val 705 710 715 aat ggt aat aat aca gaa gag etc ttt aaa gta tta aat aat ata aaa 2330 Asn Gly Asn Asn Thr Glu Glu Leu Phe Lys Val Leu Asn Asn Ile lys 720 725 730 735 gaa aat aaa tta sss aga gct act gtt ctt cat gta cgt aca aaa aaa 2378 Glu Asn Lys Leu Lys Arg Ala Thr Val Leu His Val Arg Thr Lys Lys 7 40 745 750 tcg aat gat ttt ata aat tca aag agt cca ata agt ata ttg cac tct 2426 Ser Asn. Asp Phe Ile Asn Ser Lys Ser Pro Ile Ser Ile Leu His Ser 755 760 765 ata aag aaa aat gag att ttc cct ttc gat acc act ata tta aat gga 2474 Ile Lys Lys Asn Glu Ile Phe Pro Phe Asp Thr Thr Ile Leu Asn Gly 770 775 780 aat att est aag gag aac aag ata gaa gaa gag aaa aat gtg tct tca 2522 Asn Ile His Lys Glu Asn Lys Ile Glu Glu Glu Lys Asn Val Ser Ser 785 790 795 tct aca aag tat gat gta aat aat aag aat aat aaa aat aat gat aat 2570 Ser Thr Lys Tyr Asp Val Asn Asn Lys Asn Asn Lys Asn Asn Asp Asn 800 805 810 815 agt gaa att ata àaa tat gaa gat atg ttt tca aaa gag aeg ttc aca 2618 Ser Glu Ile Ile Lys Tyr Glu Asp Met Phe Ser Lys Glu Thr Phe Thr 820 825 830 gat ata tat aca aat gaa atg tta aaa tat tta aag aaa gat aga aat 2666 Asp Ile Tyr Thr Asn Glu Met Leu Lys Tyr Leu Lys Lys Asp Arg Asn 835 840 845 ata ata ttc cta tct ccc gct atg tta gga gga tca gga ttg gtt aaa 2714 Ile Ile Phe Leu Ser Pro Ala Met Leu Gly Gly Ser Gly Leu Val Lys 850 855 860 att agt gag cgt tat cca aat aat gta tat gat gta ggt ata gea gaa 2762 Ile Ser Glu Arg Tyr Pro Asn Asn Val Tyr Asp Val Gly Ile Ala Glu 865 870 875 caa cat tct gta act ttc gea gea gct atg gea atg aat aag aaa tta 2810 Gin His Ser Val Thr Phe Ala Ala Ala Met Ala Met Asn Lys Lys Leu 880 885 890 895 - 10- 011656 aaa ata caa tta tgt ata tat tcg acc ttt tta caa aga gca tat gat 2858 Lys Ile Gin Leu Cys Ile Tyr Ser Thr Phe Leu Gin Arg Ala Tyr Asp 900 905 910 caa att ata cat gat ctt aat tta caa aat ata cct tta aag gtt ata 2906 Gin Ile Ile His Asp Leu Asn Leu Gin Asn Ile Pro Leu Lys Val Ile 915 920 925 att gga aga agt gga tta gta gga gag gat ggg gca aca cat caa ggt 2954 Ile Gly Arg Ser Gly Leu Val Gly Glu Asp Gly Ala Thr His Gin Gly 930 935 940 ata tat gat tta tct tat ctt ggg aca ctt aac aat gca tat ata ata 3002 Ile Tyr Asp Leu Ser Tyr Leu Gly Thr Leu Asn Asn Ala Tyr Ile Ile 945 950 955 tct cca agt aat caa gtt gat ttg aaa aga gct ctt agg ttt gct tat 3050 Ser Pro Ser Asn Gin Val Asp Leu Lys Arg Ala Leu Arg Phe Ala Tyr 960 965 970 975 tta gat aag gac cat tct gtg tat ata cgt ata ccc aga atg aac ata 3098 Leu Asp Lys Asp His Ser Val Tyr Ile Arg Ile Pro Arg Met Asn Ile 980 985 990 tta agt gat aag tac atg aaa gga tat ttg aac att cat atg aaa aat 3146 Leu Ser Asp Lys Tyr Met Lys Gly Tyr Leu Asn Ile His Met Lys Asn 995 1000 1005 gag agc aaa aat atc gat gta aac gtg gat ata aac gat gat gta gat 3194 Glu Ser Lys Asn Ile Asp Val Asn Val Asp Ile Asn Asp Asp Val Asp 1010 1015 1020 aaa tat agt gaa gaa tat atg gac gat gat aat ttt ata aaa tcg ttt 3242 Lys Tyr Ser Glu Glu Tyr Met Asp Asp Asp Asn Phe Ile Lys Ser Phe 1025 1030 1035 att gga aaa tct aga att att aaa atg gat aat gaa aat aat aat aca 3290 Ile Gly Lys Ser Arg Ile Ile Lys Met Asp Asn Glu Asn Asn Asn Thr 1040 1045 1050 1055 aat gaa cat tat tca agc aga gga gat aca cag aca aaa aaa aaa aaa 3338 Asn Glu His Tyr Ser Ser Arg Gly Asp Thr Gin Thr Lys Lys Lys Lys 1060 1065 1070 gtt tgt atc ttt aac atg ggt agt atg ctt ttt aat gta att aat gct 3386 Val Cys Ile Phe Asn Met Gly Ser Met Leu Phe Asn Val Ile Asn Ala 1075 1080 1085 ata aaa gaa att gaa aaa gaa caa tat att tca cat aat tat tct ttt 3434 Ile Lys Glu Ile Glu Lys Glu Gin Tyr Ile Ser His Asn Tyr Ser Phe 1090 1095 1100 tca att gtt gat atg ata ttt tta aat cct tta gat aaa aat atg ata 3482 Ser Ile Val Asp Met Ile Phe Leu Asn Pro Leu Asp Lys Asn Met Ile 1105 1110 1115 - 11 - 011656 gat catAsp His1120 gta Val ata Ile aaa Lys caa Gin 1125 aat Asn aaa Lys cat His caa Gin tat Tyr 1130 tta Leu att Ile act Thr tat Tyr gaa Glu 1135 3530 gat aat act ata ggt ggt ttt tct aca cat ttc aat aat tat tta ata 3578 Asp Asn . Thr Ile Gly 1140 Gly Phe Ser Thr His 1145 Phe Asn Asn Tyr Leu 1150 Ile gaa aat aat tat att aca aaa cat aac tta tat gtt cat aat att tat 3626 Glu Asn Asn Tyr 1155 Ile Thr Lys His Asn 1160 Leu Tyr Val His Asn 1165 Ile Tyr tta tct aat gag cca att gaa cat gca tct ttt aag gat caa caa gaa 3674 Leu Ser Asn117 0 Glu Pro Ile Glu His 1175 Ala Ser Phe. Lys Asp 1180 Gin Gin Glu gtc gtc aaa atg gat aaa tgt agt ctt gtc aat aga att aaa aat tat 3722 Val Val1185 Lys Met Asp Lys Cys 1190 Ser Leu Val Asn Arg 1195 Ile Lys Asn Tyr ctt aaaLeu Lys aat Asn aat Asn cct Pro aca Thr tgatgtaaga taaatatata t' ttctaaaat 3770 1200 1205 tatttttttt ttatacttta atgtgtacaa taaaatatat atctaaatat attttatttg 3830 tacgcttttt tttttttttt tttaattgtt atttttgtat at 3872 <210> 4 <211> 1205 <212> FRT <213> Plasmodium faiciparum <400> 4 Met Ile Fhe Asn Tyr Val Phe Phe Lys Asn Phe Val Pro Val Val Leu15 10 15 Tyr Ile Leu Leu Ile Ile Tyr Ile Asn Leu Asn Gly Met Asn Asn Lys20 25 30 Asn Gin Ile Lys Thr Glu Lys Ile Tyr Ile Lys Lys Leu Asn Arg Leu35 .40 45 Ser Arg Lys Asn Ser Leu Cys Ser Ser Lys Asn Lys Ile Ala Cys Leu50 ' 55 60 Phe Asp Ile Gly Asn Asp Asp Asn Arg Asn Thr Thr Tyr Gly Tyr Asn 65 70 75 80 Val Asn Val Lys Asn Asp Asp Ile Asn Ser Leu Leu Lys Asn Asn Tyr 85 90 95 Ser Asn Lys Leu Tyr Met Asp Lys Arg Lys Asn Ile Asn Asn Val Ile100 105 110 Ser Thr Asn Lys Ile Ser Gly Ser Ile Ser Asn Ile Cys Ser Arg Asn115 120 125 - 12- 011656 Gin Lys130 Cys His145 Asn Asn Phe Asn Phe Tyr Lys Lys210 Arg Asp225 Asn Asn Asn Asn Asp Asn Arg Ser290 Asn Asn305 Asn Thr Ile Tyr Tyr Tyr Phe Asp370 Glu Glu385 Tyr Tyr Phe Pro Glu Asn Thr Tyr Arg Asn Leu Lys180 Cys Lys195 Asn Cys Ser His Ala Leu Asn Lys260 Asn Asp27 5 Asn His Asn Asn Phe Ile Lys Glu340 Glu Arg355 Ile Asp Phe Ile Lys Lys Leu Leu 420 Glu Gin Asn Met150 Asn Lys165 Arg Met Glu Lys Thr Phe Lys Leu230 Tyr Glu245 Asn Asn Tyr Asn Tyr Asp Asp Lys310 Asn Ile325 Ile Tyr Lys Tyr Lys Tyr Asn Asn390 Glu Asn405 Lys Leu Lys Arg135 Ser His Lys Asn Lys Asn Lys Leu200 Gin Asn215 Phe Ser Ser Glu Asn Asn Asn Asn280 Asn Tyr295 Tyr Asp Asp Glu Glu Leu Phe Ser360 Asn Asp375 Gly Val Ile Leu Ile Asn Asn Lys Glu Gin Phe Asn170 Ser Leu185 Ser Phe Tyr Ser Gly Glu Lys Lys250 Lys Asn265 Asn Ser Gly Gly Ile Gly Tyr Lys330 Tyr Val345 Glu Asp Val Glu Tyr Ile Ile Met410 Asn Pro425 Gin Arg140 Asp Lys155 Leu Leu Leu Asn Leu His Leu Lys220 Phe Asp235 Glu Tyr Asn Asp Cys Asn Asp Asn300 Lys Tyr315 Thr Ile Glu Arg Ile Lys Phe Glu380 Asn Asn395 Lys Lys Ser Asp Cys Leu Leu Ala Phe Ile Lys Asp190 Lys Ala205 Arg Lys Asp Tyr Ile Thr Asn Lys270 Asn Leu285 Asn Asn Phe Lys Tyr Gly Asn Ile350 Lys Ser365 Lys Ala Ile Asp Ile Leu Leu Lys 430 Thr Gin Asn Asp160 Asn Tyr175 Asn Phe Tyr Lys Ser Asn Thr Asn240 Leu Asn255 Asn Asn Gly Glu Pro Cys Gin Ile320 Asp Glu335 Pro Glu Val Leu Ile Lys Asn Thr400 His Tyr.415 Lys Leu - 13 - 011656 Lys Lys Gin Tyr Leu Pro Leu Leu435 440 Phe Phe Ile Val Asn Ile Thr Gly450 455 Ser Leu Glu Ile Gin Leu Leu Leu465 470 Asp Asn Val Ile Tyr Asp Ile Gly485 Leu Thr Gly Arg Lys Leu Leu Phe500 Ile Ser Gly Phe Leu Asn Ile Phe515 520 Ala Gly His Ser Ser Thr Ser Leu 530 535 Ala Glu Trp Gin Val Lys Asn Lys 545 ’ 550 Glu Ile Sar Asp Asn Ala Asn Val 565 Lys Gly Ile His Asn Asp Asn Asn580 Asn Tyr Ile Asn Pro Ser Asp Val5?5 600 Pro Asn Val Arg Asn Asp Asn His 610 615 Ile Ile Gly7 Asp Gly Gly Leu Thr 625 ' 630 Asn Tyr Ile Ser Phe Leu Asn Ser 645 Asn Gly Gin Val Ser Leu Pro Thr660 Arg Pro Ile Gly Ser Ile Ser Asp675 680 Ile Glu A.ia Asn Ala Gly Asp Asn690 695 Asn Asn Ile Phe Glu Asn Leu Asn705 710 Gly Asn Asn Thr Glu Glu Leu Phe 725 Ala His Glu Leu Lys 445 Ile Phe Leu Gly His Phe Ser 460 Ser Val Leu Ser Leu Tyr Ile 475 Phe Asn Gin Pro Tyr 480 His Gin 490 Ala Tyr Val His Lys 495 Ile Leu 505 Ser Leu Arg Asn Lys 5.10 Lys Gly Glu Ser Ile Tyr Asp525 Lys Phe Gly Ser Ala Ile Gin 540 Gly Tyr Tyr Glu Glu Lys Tyr 555 Gly Asn Gly Asp Ile 560 Thr Asn 570 Asn Glu Arg Ile Phe 575 Gin Ile 585 Asn Asn Asn Ile Asn 590 Asn Asn Val Gly Arg Glu Asn 605 Thr Asn Val Asn Val Asp Lys 620 Val His Ile Ala Gly Gly Met 635 Ala Leu Glu Ala Leu 640 Lys Ile 650 Leu Ile Ile Tyr Asn 655 Asp Asn 665 Ala Val Ser Ile Ser 670 Gly Asn His Leu His Tyr Phe 685 Val Ser Asn Lys Leu Ser Lys 700 Asn Ala Lys Glu Tyr Asp Tyr 715 Ile Gly Val Val Asn 720 Lys Val 730 Leu Asn Asn Ile Lys 735 Glu - 14- 011656 Asn Lys Leu Lys Arg Ala740 Thr Val Leu His745 Val Arg Thr Lys750 Lys Ser Asn Asp Phe Ile Asn Ser Lys Ser Pro Ile Ser Ile Leu His Ser Ile 755 760 765 Lys Lys Asn Glu Ile Phe Pro Phe Asp Thr Thr Ile Leu Asn Gly Asn 770 775 780 Ile His Lys Glu Asn Lys Ile Glu Glu Glu Lys Asn Var Ser Ser Ser 785 790 795 800 Thr Lys Tyr Asp Val Asn Asn Lys Asn Asn Lys Asn Asn Asp Asn Ser 805 810 815 Glu Ile Ile Lys Tyr Glu Asp Met Phe Ser Lys Glu Thr Phe Thr Asp 820 825 830 Ile Tyr Thr Asn Glu Met Leu Lys Tyr Leu Lys Lys Asp Arg Asn Ile 835 840 845 Ile Phe Leu Ser Pro Ala Met Leu Gly Gly Ser Gly Leu Val Lys Ile 850 855 860 Ser Glu Arg Tyr Pro Asn Asn Val Tyr Asp Val Gly Ile Ala Glu Gin 865 870 875 880 His Ser Val Thr Phe Ala Ala Ala Met Ala Met Asn Lys Lys Leu Lys 885 890 895 Ile Gin Leu Cys Ile Tyr Ser Thr Phe Leu Gin Arg Ala Tyr Asp Gin 900 905 910 Ile Ile His Asp Leu Asn Leu Gin Asn Ile Pro Leu Lys Val Ile Ile 915 920 925 Gly Arg Ser Gly Leu Val Gly Glu Asp Gly Ala Thr His Gin Gly Ile 930 935 940 Tyr Asp Leu Ser Tyr Leu Gly Thr Leu Asn Asn Ala Tyr Ile Ile Ser 945 950 955 960 Pro Ser Asn Gin Val Asp Leu Lys Arg Ala Leu Arg Phe Ala Tyr Leu 965 970 975 Asp Lys Asp His Ser Val Tyr Ile Arg Ile Pro Arg Met Asn Ile Leu 980 985 990 Ser Asp Lys Tyr Met Lys Gly Tyr Leu Asn Ile His Met Lys Asn Glu 995 1000 1005 Ser Lys Asn Ile Asp Val Asn Val Asp Ile Asn Asp Asp Val Asp Lys 1010 1015 1020 Tyr Ser Glu Glu Tyr Met Asp Asp Asp Asn Phe Ile Lys Ser Phe Ile 025 1030 1035 1040 - 15- 011656 Gly Lys Ser Arg Ile 1045 Ile Lys Met Asp Asn 1050 Glu Asn Asn Asn Thr 1055 Asn Glu His Tyr Ser1060 Ser Arg Gly Asp Thr 1065 Gin Thr Lys Lys Lys 1070 Lys Val Cys Ile Phe Asn1075 Met Gly Ser Met1080 Leu Phe Asn Val Ile 1085 Asn Ala Ile Lys Glu 1090 Ile Glu Lys Glu Gin Tyr1095 Ile Ser His Asn 1100 Tyr Ser Phe Ser Ile 105 Val Asp Met Ile Phe 1110 Leu Asn Pro Leu Asp 1115 Lys Asn Met Ile Asp 1120 His Val Ile Lys Gin 1125 Asn Lys His Gin Tyr 1130 Leu Ile Thr Tyr Glu 1135 Asp Asn Thr Ile Gly1140 Gly Phe Ser Thr His 1145 Phe Asn Asn Tyr Leu 1150 Ile Glu Asn Asn Tyr Ile1155 Thr Lys His Asn1160 Leu Tyr Val His Asn 1165 Ile Tyr Leu Ser Asn 1170 Glu Pro Ile Glu His Ala1175 Ser Phe Lys Asp 1180 Gin Gin Glu. Val Val 185 Lys Met Asp Lys Cys 1190 Ser Leu Val Asn Arg 1195 Ile Lys Asn Tyr Leu 1200 Lvs Asn Asn Pro Thr 1205 <210> 5 <211> 3147 <212> DNA <213> Plasmodium falciparum <220> <221> CDS<222> (199: !..(2670) <400> 5 tttcattttt ctttacccac atatatatat tatatttgat atatgattta aaattgraac atacatctcc aacatataaa tattatrttt atatatatat aatatatata tataatatta 60 ataaaaaaaa taattatatt aaatatgtgt 120 tattattatt tttttttttt tttttcataa 180 231 tgcctgaata accacaaa atg Met 1 agt Ser tat Tyr ata Ile aaa Lys 5 aga Arg ctg Leu att Ile ctt Leu ttt Phe 10 atg Met tta ctg ttt tat tct cat gta aaa att aaa aaa tta ttt att aaa att Leu Leu Phe Tyr Ser His Val Lys Ile Lys Lys Leu Phe Ile Lys Ile 15 20 25 - 279 - 16- 011656 tct Ser aat Asn Vs _ aac Asn ata Ile ttt Phe ttt Phe gcaAl a 55 gaa Glu gca Ala aag Lys aaa Lys aat Asn 40 gga Gly aaa Lys aag Lys 327 gaa ttc tt~ ctt ttt tta cta aat ata aaa aaa aat agc caa cag aaa 375 Glu Phe Phe Leu Phe Leu Leu Asn Ile Lys Lys Asn Ser Gin Gin Lys 45 50 55 33â act t S Z cat att acc aaa agg aat acc ata aat aaa agt gat ttt 423 Lys Thr Tyr His Ile Thr Lys Arg Asn Thr Ile Asn Lys Ser Asp Phe 60 65 70 75 tta tat tct tta cta aat gaa gaa ggg aat tct tca aaa aag gaa tat 471 Leu Tyr Ser Leu Leu Asn Glu Glu Gly Àsn Ser Ser Lys Lys Glu Tyr 80 85 90 aaa aat tta aaa gat gaa gaa 33 3 tat aat atc ata caa aat ata aaa 519 Lys Asn Leu Lys Asp Glu Glu Lys Tyr Asn Ile Ile Gin Asn Ile Lys 95 100 105 aaa tat tgt gaa tgt act aaa 333 tat aaa agg etc cca aca ega gaa 567 Lys Tyr Cys Glu Cys Thr Lys Lys Tyr Lys Arg Leu Pro Thr Arg Glu 11 1 115 120 gta gtt s-î gga aat gtt aaa att gga gga aat aat aaa ata gct att 615 Val Val i _ e Gly Asn Val Lys Ile Gly Gly Asn Asn Lys Ile Ala Ile 125 130 135 caa act. S ~ 3 gct agc tgt gat SCS aga aat gta gaa gaa tgt gta tat 663 Gin Thr Mec Ala Ser Cys Asp Thr Arg Asn Val Glu Glu Cys Val Tyr 14 0 145 150 155 caa atc ara aaa tgt aaa gat ttg ggt gct gac att gta agg ttg act 711 Gin Ile Ar ; Lys Cys Lys Asp Leu Gly Ala Asp Ile Val Arg Leu Thr 160 165 170 gtt caa a c a gtt caa gaa gca caa gct agt tat cat att aaa gaa aaa 759 Val Gin Gl·.· Val Gin Glu Ala C-ln Ala Ser Tyr His Ile Lys Glu Lys 175 180 185 tta tta tct gaa aat gta aat cüC cca tta gta gca gat att cat ttt 807 Leu Leu Ser Glu Asn Val Asn. Ile Pro Leu Val Ala Asp Ile His Phe 191 195 200 S 31 cct 3 S S ata gct tta atg gca gct gat gtg ttt gaa aaa att ega 855 Asn Pro Lys Ile Ala Leu Met Ala Ala Asp Val Phe Glu Lys Ile Arg 205 210 215 gtg aat cca gga aat tat gtt gat gga aga aaa aaa tgg ata gat aaa 903 Val Asn Prc Gly Asn Tyr Val Asp Gly Arg Lys Lys Trp Ile Asp Lys 220 225 230 235 gtt tat 333 act aaa gaa gaa ttt gat gaa ggg aaa tta ttt ata aaa 951 Val Tyr Lys Thr Lys Glu Glu Phe Asp Glu Gly Lys Leu Phe Ile Lys 240 245 250 - 17- 011656 gaa Glu aaa Lys ttt Phe gta Val 255 CCS Pro tta Leu att Ile gaa Glu aaa Lys 260 tgt Cys aaa Lys aga Arg tta Leu aat Asn 265 aga Arg gcaAl a 999 ata aga at t gga aca aat cat gga tcc ctt tca tct cga gta tta tca 1047 Ile Arg Ile Gly Thr Asn His Gly Ser Leu Ser Ser Arg Val Leu Ser 27 0 275 280 tat tat gga gat aca cca tta ggt atg gta gaa tcg gct ttt gag ttt 1095 Tyr Tyr Gly Asp Thr Pro Leu Gly Met Val Glu Ser Al a Phe Glu Phe 285 290 295 tct gat tt a tgt att gaa aac aat ttt tac aat ctt gtt ttt tct atg 1143 Ser Asp Leu Cvs lie Glu A.sn Asn Phe Tyr Asn Leu Val Phe Ser Met 300 305 310 315 aaa gct tct Sat cct tat gtt àtg —^-4* —.O uo caa tct tat aga tta tta gta 1191 Lys Al a Ser Asn Al a Tyr Val Met Ile Gin Ser Tyr Arg Leu Leu Val 320 325 330 tct aaa CSS tat çaa aga aat atg atg ttc cct ata cat tta gga gtt 1239 Ser Lys Gin Tyr Glu Arg Asn Met Met Phe Pro Ile His Leu Gly Val - 335 340 345 aca gaa gca gga ttt çgt Ç31 aat gga aga ata aaa tct tat tta ggt 1287 Thr Glu Al a Gly Phe GÎy Asp Asn Gly Arg Ile Lys Ser Tyr Leu Gly 350 355 360 ata gga tct tta tta tat gat ggt ata gga gat acc att cgt ata tcc 1335 Ile Gly Ser Leu Leu Tyr Asp Gly Ile Gly Asp Thr Ile Arg Ile Ser 365 370 375 tta aca gaa gat cct tgg gaa gag tta act cct tgt 333 aaa tta gtt 1383 Leu Thr Glu Asp Pro Trp Glu Glu Leu Thr Pro Cys Lys Lys Leu Val 380 385 390 395 gaa aat tta aag aaa aga ata ttt tat aat gaa aat ttt aaa gaa gat 1431 Glu Asn Leu Lys Lys Arg Ile Phe Tyr Asn Glu Asn Phe Lys Glu Asp 400 405 410 aat gaa tta aaa aat aat gaa atg gat acc aaa aat cta tta aat ttt 1479 Asn Glu Leu Lys Asn Asn Glu Met Asp Thr Lys Asn Leu Leu Asn Phe 415 420 425 gaa gaa 3ât tat cga aat ttt 5.31 aat ata aaa aaa aga aat gta gaa 1527 Glu Glu Asn Tyr Arg Asn Phe Asn Asn Ile Lys Lys Arg Asn Val Glu 430 435 440 aaa aat aat aat gta tta cat 033 gag tgc act ata ggt aat gta gta 1575 Lys Asn Asn Asn Val Leu His Glu Glu Cys Thr Ile Gly Asn Val Val 445 450 455 acc ata aaa gag tta gaa gat tct ctg caa att ttt aaa gat tta aat 1623 Thr Ile Lys Glu Leu Glu Asp Ser Leu Gin Ile Phe Lys Asp Leu Asn 460 465 470 475 - 18 - 011656 tta Leu gaa Glu gta Val gat Asp tca Ser 480 aat Asn gga Gly aat Asn ttg Leu aaa Lys 485 aag Lys gga G-ly gcc Ala aaa Lys aca Thr 490 act Thr 1671 gat atg gtt att ata aat gat ttt cat aat ata aca aat tta gga aaa 1719 Asp Met Val Ile Ile Asn Asp Phe His Asn Ile Thr Asn Leu Gly Lys 495 500 505 aaa act gtg gat aaa tta atg caa gtg gga att aat ata gta gtt caa 1767 Lys Thr Val Asp Lys Leu Met Gin Val Gly Ile Asn Ile Val Val Gin 510 515 520 tat gaa cca cat aat ata gaa ttt ata. gaa 533 atg gaa cca aat aat 1815 Tyr Glu Pro His Asn Ile Glu Phe Ile Glu Lys Met Glu Pro Asn Asn 525 530 535 gat 331 aat aat aat aat aat aat aat aat ata tta ttt tat gtg gat 1863 Asp Asn Asn Asn Asn Asn Asn Asn Asn Asn Ile Leu Phe Tyr Val Asp 540 545 550 555 ata aaa 33 l att atg aac agt tca gaa aaa aat att aaa tta agt aat 1911 Ile Lys Asn Ile Met Asn Ser Ser Glu Lys Asn Ile Lys Leu Ser Asn 560 565 570 tct aaa gga tat gga tta att tta aac gga aaa gaa gat ata caa acc 1959 Ser Lys Gly Tyr Gly Leu Ile Leu Asn Gly Lys Glu Asp Ile Gin Thr 575 580 585 ata aaa asa ata aaa gaa tta aat cgt cgt cct tta ttc att cta tta 2007 Ile Lys Lys Ile Lys Glu Leu Asn Arg Arg Pro Leu Phe Ile Leu Leu 5 90 595 600 aaa tca gat aac ata tat gaa cat gta tta ata acc aga aga att aat 2055 Lys Ser Asp Asn Ile Tyr Glu His Val Leu Ile Thr Arg Arg Ile Asn 605 610 615 gaa ctt tta caa tcc tta aat ata aat ata cct tat ata cat tat gtt 2103 Glu Leu Leu Gin Ser Leu Asn Ile Asn Ile Pro Tyr Ile His Tyr Val 620 625 630 635 gat att aat tca aac aat tat gat gat ata tta gtt aat tca aca tta 2151 Asp Ile Asn Ser Asn Asn Tyr Asp Asp Ile Leu Val Asn Ser Thr Leu 640 645 650 tat gca gga agt tgt ttg atg gat tta atg ggg gat ggt ctt att gtt 2199 Tyr Al a Gly Ser Cys Leu Met Asp Leu Met Gly Asp Gly Leu Ile Val 655 660 665 aac gta oCtL aat gat gtt ctt 5.C3 aat aaa aaa aag ata gaa aca aaa 2247 Asn Val Thr Asn Asp Val Leu Thr Asn Lys Lys Lys Ile Glu Thr Lys 670 675 680 tat gat gaa aaa gaa gaa gta gag gaa gag gga aac aat aaa gat att 2295 Tyr Asp Glu Lys Glu Glu Val Glu Glu Glu Gly Asn Asn Lys Asp Ile 685 690 695 - 19- 011656 cat His 700 aga Arg et t Leu ttg Leu age Ser aga Arg 705 gtt Val gea Al a tta Leu aat Asn tca Ser 710 ttt Phe t U. 3. Leu aca Thr tta Leu aat Asn 715 2343 att tta caa gat aca aga ata cgt tta ttt aaa SCS gat tat ata gcc 2391 Ile Leu Gin Asp Thr Arg Ile Arg Leu Phe Lys Thr Asp Tyr Ile Ala 720 725 730 tgc cca tet tgt gga aga act tta ttt aat ata caa taa act 2 Ctl aaa 2439 Cys F'ro Ser Cys Gly Arg Thr Leu Phe Asn Ile Gin Glu Thr Thr Lys 735 740 745 άάά att atg aaa tta aca ggg cac tta aaa ggc gtt âââ att gea gtc 2487 Lys Ile Met Lys Leu Thr Gly Kis Leu Lys Gly Val Lys Ile Ala Val 750 n55 7 60 atg aga tgt att gtt aat ggt ara gga gaer atg gea gat gea cat ttt 2535 Met Gly Cvs Ile Val Asn Gly Ile Giv Glu Met Al a Asp Al a His Phe 765 770 775 ggt tat gtt ggt agt gea cct aaa aaa att gat tta tat tat ggt aaa 2583 Gly Tyr Val Gly Ser Ala Pro Lys Lys Ile Asp Leu Tyr Tyr Gly Lys 780 785 7 90 795 gag tta gta gaa aga aat ata cct gag gaa gaa gct tgt gat aaa ttg 2631 Glu Leu Val Glu Arg Asn Ile Fro Glu Glu Glu Al a Cys Asp Lys Leu 800 805 810 ata gaa tta att aaa aaa cat sac aaa tgg aaa gat cca taaattgaat 2680 Ile Glu Leu Ile Lys Lys His Asn Lys Trp Lys Asp Pro 815 820 atggacaagt atttatttat ttatttatct tatatataat atattataaa tttttcgatg 2740 tatttttcct tttaaaattt tatttttttt ttattttttt ttttgaagta atatttataa 2800 tgcatacata atattaaaat gtgtattata taataatatc attttattgt tattttaaaa 2860 gactaatacc aagaacaatt ttttaataat cattcttata acttgttaaa tatatatata 2920 tatatatata tatttattta tttatartta tatttattta tttttggtat atgaaaagta 2980 aaaatataat aatttaaaag tatttacaaa ataaataata ttatatatct gtttttatat 3040 atatgttaat ggaaaaggag aaaataaata aataaaacaa acaaaataac atatatatat 3100 atatatatat actgaatgag aaagaaaaaa aaaagaaaag gatacga 3147 <210> 6 <2U> 824 <212> PRT <213> Plasmodium falciparum <400> 6 Met Ser Tvr Ile Lys Arg Leu Ile Leu Phe Met Leu Leu Phe Tyr Ser 1 ’ 5 10 15 -20- 011656 His Val Lys Ile Lys Lys Leu Phe Ile Lys Ile Ser Asn Val Asn Ile20 25 30 Phe Phe Ala Glu Ala Lys Lys Asn Gly Lys Lys Glu Phe Phe Leu Phe35 40 45 Leu Leu Asn Ile Lys Lys Asn Ser Gin Gin Lys Lys Thr Tyr His Ile50 55 60 Thr Lys Arg Asn Thr Ile Asn Lys Ser Asp Phe Leu Tyr Ser Leu Leu65 70 75 80 Asn Glu Glu Gly Asn Ser Ser Lys Lys Glu Tyr Lys Asn Leu Lys Asp85 90 95 Glu Glu Lys Tyr Asn Ile Ile Gin Asn Ile Lys Lys Tyr Cys Glu Cys100 105 ' 110 Thr Lys Lys Tyr Lys Arg Leu Pro Thr Arg Glu Val Val Ile Gly Asn115 120 125 Val Lys Ile Gly Gly Asn Asn Lys Ile Ala Ile Gin Thr Met Ala Ser130 135 140 Cys Asp Thr Arg Asn Val Glu Glu Cys Val Tyr Gin Ile Arg Lys Cys 145 150 155 160 Lys Asp Leu Gly Ala Asp Ile Val Arg Leu Thr Val Gin Gly Val Gin 165 170 175 Glu Ala Gin Ala Ser Tyr His Ile Lys Glu Lys Leu Leu Ser Glu Asn180 185 190 Val Asn Ile Pro Leu Val Ala Asp Ile His Phe Asn Pro Lys Ile Ala195 200 205 Leu Met Ala Ala Asp Val Phe Glu Lys Ile Arg Val Asn Pro Gly Asn210 215 220 Tyr Val Asp Gly Arg Lys Lys Trp Ile Asp Lys Val Tyr Lys Thr Lys 225 230 235 240 Glu Glu Phe Asp Glu Gly Lys Leu Phe Ile Lys Glu Lys Phe Val Pro 245 250 255 Leu Ile Glu Lys Cys Lys Arg Leu Asn Arg Ala Ile Arg Ile Gly Thr 260 " 265 270 Asn His Gly Ser Leu Ser Ser Arg Val Leu Ser Tyr Tyr Gly Asp Thr 275 280 285 Pro Leu Gly Met Val Glu Ser Ala Phe Glu Phe Ser Asp Leu Cys Ile 290 " 295 300 Glu Asn Asn Phe Tyr Asn Leu Val Phe Ser Met Lys Ala Ser Asn Ala 305 310 315 320 Tyr Val Met Ile Gin Ser Tyr Arg Leu Leu Val Ser Lys Gin Tyr Glu -21 - 011656 325 Arg Asn Met Met rte Pro Ile His340 Gly Asp Asn Gly Arg Ile Lys Ser355 360 Tyr Asp Gly Ile Gly Asp Thr Ile370 375 Trp Glu Glu Leu Thr Pro Cys Lys385 390 Arg Ile Phe Tyr Asn Glu Asn Phe405 Asn Glu Met Asp Thr Lys Asn Leu420 Asn Phe Asn Asn Ile Lys Lys Arg435 440 Leu His Glu Glu Cys Thr Ile Gly450 455 Glu Asp Ser Leu Gin Ile Phe Lys465 470 Asn Gly Asn Leu Lys Lys Gly Ala485 Asn Asp Phe His Asn Ile Thr Asn500 Leu Met Gin Val Gly Ile Asn lie515 520 Ile Glu Phe Ile Glu Lys Met Glu530 535 Asn Asn Asn Asn Asn Ile Leu Phe545 550 Asn Ser Ser Glu Lys Asn Ile Lys565 Leu Ile Leu Asn Gly Lys Glu Asp580 Glu Leu Asn Arg Arg Pro Leu Phe595 600 Tyr Glu His Val Leu Ile Thr Arg610 615 Leu Asn Ile Asn Ile Pro Tyr Ile 625 630 330 335 Leu Gly Val Thr Glu Ala Gly Phe345 350 Tyr Leu Gly Ile Gly Ser Leu Leu365 Arg Ile Ser Leu Thr Glu Asp Pro380 Lys Leu Val Glu Asn Leu Lys Lys395 400 Lys Glu Asp Asn Glu Leu Lys Asn410 415 Leu Asn Phe Glu Glu Asn Tyr Arg425 430 Asn Val Glu Lys Asn Asn Asn Val445 Asn Val Val Thr Ile Lys Glu Leu460 Asp Leu Asn Leu Glu Val Asp Ser475 480 Lys Thr Thr Asp Met Val Ile Ile490 495 Leu Gly Lys Lys Thr Val Asp Lys505 510 Val Val Gin Tyr Glu Pro His Asn525 Pro Asn Asn Asp Asn Asn Asn Asn540 Tyr Val Asp Ile Lys Asn Ile Met555 560 Leu Ser Asn Ser Lys Gly Tyr Gly570 575 Ile Gin Thr Ile Lys Lys Ile Lys585 590 Ile Leu Leu Lys Ser Asp Asn Ile605 Arg Ile Asn Glu Leu Leu Gin Ser620 His Tyr Val Asp Ile Asn Ser Asn635 6*4 0 -22- 011656 Asn Tyr Asp Asp Ile Leu Val Asn Ser Thr Leu Tyr Ala Gly Ser Cys645 650 655 Leu Met Asp Leu Met Gly Asp Gly Leu Ile Val Asn Val Thr Asn Asp 660 ' 665 670 Val Leu Thr Asn Lys Lys Lys Ile Glu Thr Lys Tyr Asp Glu Lys Glu 675 680 685 Glu Val Glu Glu Glu Gly Asn A.sn Lys Asp Ile His Arg Leu Leu Ser690 695 700 Arg Val Ala Leu Asn Ser Phe Leu Thr Leu Asn Ile Leu Gin Asp Thr 705 710 715 720 Arg Ile Arg Leu Phe Lys Thr Asp Tyr Ile Ala Cys Pro Ser Cys Gly 725 730 735 Arg Thr Leu Phe Asn Ile Gin Glu Thr Thr Lys Lys Ile Met Lys Leu740 745 750 Thr Gly His Leu Lys Gly Val Lys Ile Ala Val Met Gly Cys Ile Val755 760 765 Asn Gly Ile Gly Glu Met Ala Asp Ala His Phe Gly Tyr Val Gly Ser770 775 780 Ala Pro Lys Lys Ile Asp Leu Tyr Tyr Gly Lys Glu Leu Val Glu Arg 785 790 795 800 Asn Ile Pro Glu Glu Glu Ala Cys Asp Lys Leu Ile Glu -Leu Ile Lys 805 " 810 815 Lys His Asn Lys Trp Lys Asp Pro820