TH12985C3 - Plasmid carriers for the production of targeted proteins outside of bacterial cells. Lactobacillus plantarum and Bacillus subtilis. - Google Patents

Abstract

This invention-based carrier plasmid is a plasmid that can be used to produce target proteins. Extracellular in host bacteria recognized for their Generally Recognized As Safe (GRAS) from the FDA (US Food and Drug Administration), popularly used in genetic work. Currently engineered for extracellular target protein, both Lactobacillus plantarum and Bacillus subtilis provide new applications. Which the plasmid carrier according to this invention only The single structure offers extensive applications for the two bacteria, and the fabricated plasmid composition facilitates the production of target proteins outside the bacterial cells, but Each type effectively Thus making it convenient and easy to apply target proteins. Reduce the extraction process Protein is removed from the cell as compared to the target protein production used Enable to reduce Time and production costs can be reduced By plasmid carriers according to this invention Characteristically, the plasmid carriers contain a component of the protein-building blocks involved in the extracellular protein (signal peptide), which was isolated from L. plantarum BCC9546 and the aforementioned sequence. Can be selected from either the baseline sequence shown in Figure 1a, 1b or 1c

Claims (6)

1. Plasmid carriers for the production of targeted proteins outside the bacterial cells. Characteristically, Lactobacillus plantarum and Bacillus subtilis are those plasmids with sequencing components. Produce proteins involved in the extracellular protein bacteria, which are isolated from bacteria. Lactobacillus plantarum BCC9546 and the aforementioned sequences. You can choose from either the baseline sequence shown in Figure 1a, 1b or 1c. 2. Plasmid carriers according to claim 1, where such plasmid carriers comprise the main production baseline. The proteins are extracellular as follows: A. Origin of replication (ORI) and protein-forming bases. It was involved in the replication of plasmids (Replication protein, Rep 2) in gram-positive bacteria, which were isolated. From natural plasmids of L.plantarum bacteria BCC9546 b. Plasmid sequencing in gram-negative bacteria derived from plasmid pUC18 C. Promoter part sequencing isolated from enzyme-producing genes. L-lactate dehydrogenase (L-lactate dehydrogenase) of L.plantarum BCC9546 bacteria, whose base sequence is shown in Figure 2 d. Protein-generating base sequence involved in the extracellular protein. Isolated from either L.plantarum BCC9546 bacteria, which was selected from the sequence shown in Figure 1a, 1b or 1c, e. The sequence of the three cloning enzymes (Multiple cloning site, MCS) was EcoRl, Kpnl Smal f. Base sequencing produces an antibiotic-resistant protein, erythromycin. (Erythromycin) 3. Plasmid carriers according to claim 1 or 2, where such plasmid carriers have all bases. Can be selected from the base sequence as shown in Figure 4a, 4b or 4c, either 4. Plasmid carrier according to one of the 1-3 claims Where it is connected to the target gene 5. Selected host bacteria from L.plantarum or B.subtilis, where there are plasmid carriers. According to any 1-4 claim 6. The use of host bacteria with plasmid carriers. According to claim 5, the proliferation of target genes or the expression of target proteins to extracellate the target protein.