JPH022891A - Production of film sensitive selectively to adenine compound - Google Patents

Abstract

PURPOSE:To produce a film having selective sensitivity to compds. of adenine as a specified base in nucleic acid by forming 1-octadecylthymine into a very thin film by the Langmuir-Blodgett's technique. CONSTITUTION:1-Octadecylthymine represented by the formula is formed into a very thin film by the Langmuir-Blodgett's(LB) technique. A film having selective sensitivity to compds. of adenine as a specified base in nucleic acid can be produced.

Description

[Detailed Description of the Invention] [Summary] A method for producing a membrane selectively sensitive to a specific nucleobase, particularly a nucleobase moiety consisting of four types of compounds: adenine, thymine, guanine, and cytosine, selectively with adenine derivatives. Regarding a method for producing an interacting film, the purpose of this study is to provide a method for producing an ultra-thin film that is sensitive to adenine compounds, using the Langmuir-Blodgett (LB) method. -It consists of making octadecylthymine into an ultra-thin film.

[Industrial application field]

The present invention relates to a method for producing a membrane selectively sensitive to specific nucleobases, and particularly relates to a method for producing a membrane that selectively interacts with adenine derivatives among nucleobase moieties consisting of four types of compounds: adenine, thymine, guanine, and cytosine. .

[Background Art and Problems to be Solved] Various biosensors have been developed in recent years and are used to measure biological substances. Generally, to create a biosensor, various enzymes are immobilized on an electrode, and changes in dissolved oxygen concentration due to enzyme reactions and p
The target substance is quantified by detecting changes in l+ with electrodes. Enzymes have extremely high substrate specificity, and biosensors can selectively detect only the target substance, but enzymes are unstable because they are derived from living organisms.
Also - very expensive for boat fishing. In addition, in order to electrically detect enzyme reactions, dissolved oxygen concentration and pH
However, it can be said that it is a special case that such a condition is satisfied.

On the other hand, the development of biotechnology in recent years has been remarkable, and as a result, DNA, which is the main body of genes, is one of the substances whose quantity is currently attracting attention. The substances that make up DNA can be roughly divided into deoxyribose, phosphoric acid, and nucleobase moieties. Here, the nucleobase part is the part that controls genetic information, and there are four types of compounds: adenine, thymine, guanine, and cytosine. In particular, adenine and its related compounds are deeply involved in energy metabolism in living organisms and are the most important nucleobases.In the field of biotechnology, the determination of this adenine-related compound, especially its selection in a mixture of other similar compounds quantitative determination is extremely important.

However, although various enzymes using adenine-related compounds as substrates are known, they suffer from all of the above-mentioned problems in biosensor production and have not been put to practical use. Furthermore, if we can develop a material that selectively interacts with adenine derivatives without using enzymes and detect the interaction by some means, it will be possible to detect adenine derivatives.

However, it is extremely difficult to impart substrate specificity like enzymes to artificial substances, and this method has not yet been put to practical use.

The present invention provides super 7J cells sensitive to adenine compounds.
The purpose of the present invention is to provide a method for producing an i-film.

[Means to solve the problem]

According to the present invention, the above problem can be solved by
The problem is solved by a method for producing a membrane selectively sensitive to adenine compounds, which is characterized by forming an ultrathin film of 1-octadecylthymine represented by the following formula (I) using the (LB) method.

Unless the thickness is made extremely thin and the focal density of the thymine portion is increased, it is difficult to efficiently cause or detect interactions. In the present invention, the film is formed using the LB method, so the film thickness is at the level of 27 to 1,350 people, and the thymine portion is packed in a high density and well-ordered manner, achieving extremely efficient selective interaction. be done.

[For production]

It is known that four types of nucleobases have selective hydrogen bonds involved in genetic information transmission. In other words, adenine is thymine,
Guanine selectively hydrogen bonds with cytosine. According to the present invention, by forming a Langmuir-Blodgett (LB) film using 1-octadecylthymine represented by formula (I), it is possible to develop a material that selectively interacts with adenine-related compounds. Become. Since the above-mentioned membrane has a thymine moiety, it is possible to selectively interact with adenine in a mixed solution. Here, since hydrogen bonding is originally a weak interaction, the method for synthesizing the membrane [Example] Compound 1 is shown below.

16.4 g (0.13 wole) of thymine and 33.4 g (0.1 mole) of octadecyl bromide
After dissolving in 0 ml of dimethylformamide, 13.8
Disperse 11 g (0.1 mole) of potassium carbonate.
The reaction was carried out at 0°C for 20 hours. The reaction solution was concentrated to dryness, extracted with 150 ml of benzene, and 200 ml of n
-Hexane was added and the mixture was allowed to stand overnight at 20°C, and white crystals were precipitated. The white powder obtained by filtering this was purified by liquid chromatography to obtain 21.6 g of 1-octadecylthymine.

A 2.87×10 −3 M chloroform solution of 1-octadecylthymine was prepared, and this solution was gently dropped onto a water surface filled with pure water to form a monomolecular film of 1-octadecylthymine on the water surface. By transferring this monomolecular film onto a substrate using the LB method (surface pressure 30 mN/m), l-
Ultrathin films of octadecylthymine have been fabricated.

The selective sensitivity of membranes to adenine-related compounds was confirmed as follows. The monomolecular film prepared on the water surface as described above was used as an ion-functional field effect transistor.
(ISFET), and the gate portion of 15FET was coated with the film of the present invention. When this 15FET was immersed in phosphate buffer solutions of adenosine prepared at various concentrations, the gate current changed depending on the concentration of adenosine, confirming that the membrane was sensitive to adenine-related compounds. In addition to adenosine, cytidine,
Although guanidine and the like were added, the gate current showed a value corresponding only to the concentration of adenosine, confirming that this membrane has high selectivity for adenine-related compounds.

The properties of this membrane did not change even after being immersed in a phosphate buffer solution for several days and it was confirmed that the membrane was stable.

〔Effect of the invention〕

As explained above, according to the present invention, by producing an ultra-thin film of l-octadecylthymine by the Langmuir-Blodgett (LB) method, a film that is selectively sensitive to specific nucleobases and adenine compounds is produced. It becomes possible to create

Claims (1)

[Claims] The following (
I) A method for producing a membrane selectively sensitive to adenine compounds, which comprises forming an ultra-thin film of 1-octadecylthymine represented by the formula. ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) (CH_2)_1_7CH_3