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Nucleic acid molecules, expression vectors and applications, antibacterial drugs and methods

A nucleic acid molecule and expression vector technology, which is used in antibacterial drugs, nucleic acid molecules, expression vectors and application fields, and can solve problems such as rare reports on histone genes.

Active Publication Date: 2020-06-16
INST OF AQUATIC LIFE ACAD SINICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many reports on the role of histone variants in development and gene transcription regulation, histone-derived antimicrobial peptides and their biological functions; but the role of histone gene polymorphisms in pathogenic infection is still rarely reported

Method used

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  • Nucleic acid molecules, expression vectors and applications, antibacterial drugs and methods
  • Nucleic acid molecules, expression vectors and applications, antibacterial drugs and methods
  • Nucleic acid molecules, expression vectors and applications, antibacterial drugs and methods

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] In the process of studying the histone H2A of zebrafish, the inventors found that there are abundant nucleotide polymorphisms in the gene of the protein. The inventors further isolated four variant nucleic acid sequences of histone H2A, and named them H2A-1, H2A-2, H2A-3, and H2A-4, respectively. Sequence comparative analysis shows that the sequences of the four variant nucleic acid molecules are respectively shown in SEQ ID NO.1-4. There are a total of 42 base differences in these 4 variant nucleic acid sequences ( figure 1 ). The identity between the nucleic acid sequences of these four variants is between 91.6% and 94.9% ( figure 2 ). Among them, the amino acid sequences of the histones encoded by H2A-1, H2A-3, and H2A-4 are consistent, as shown in SEQ ID NO.5, but the amino acid sequences of these three are the same as those of the histones encoded by H2A-2 (such as SEQ ID NO.6) sequence is inconsistent, reflected in the 17th and 42nd amino acid residue types a...

experiment example 1

[0043] Effect of overexpression of the above-mentioned variant nucleic acid sequences on the proliferation of Edwardsiella caturis

[0044] In this experiment example, the influence of the overexpression of H2A-1, H2A-2, H2A-3 and H2A-4 in fish on the proliferation of Edwardsiella caturis was tested by the plate counting method. The specific operations are as follows:

[0045] 1.1 By microinjection, empty plasmids and plasmids that can express H2A-1, H2A-2, H2A-3 and H2A-4 respectively (see Figure 7 ) were microinjected into zebrafish embryos respectively;

[0046] Insert the complete open reading frames of H2A-1, H2A-2, H2A-3 and H2A-4 into the eukaryotic expression vector p3XFLAG-CMV TM -14 in.

[0047] 1.2 On the 4th day after fertilization, after the zebrafish larvae hatched, the empty plasmid group, H2A-1, H2A-2, H2A-3 and H2A-4 plasmid groups were divided into groups for soaking infection, catfish Edwards Bacterial infection concentration is 2×10 8 pfu / ml;

[0048]...

experiment example 2

[0053] Effects of in vivo overexpression of the above-mentioned variant nucleic acid sequences on the survival rate of fish infected with Edwardsiella caturis

[0054] In this experiment example, the influence of the overexpression of the three variant nucleic acid molecules of H2A-1, H2A-2 and H2A-3 in zebrafish on the survival rate under the infection of Edwardsiella catguis was tested by infection survival curve. The specific operation is as follows:

[0055] 1.1 Microinject empty plasmid FLAG, plasmids expressing H2A-1, H2A-2 and H2A-3 into zebrafish embryos by microinjection;

[0056] 1.2 On the 4th day after fertilization, after the zebrafish larvae hatched, the empty plasmid group, H2A-1, H2A-2 and H2A-3 plasmid groups were divided into groups for soaking infection, and the infection concentration was 2×10 8 pfu / ml;

[0057] 1.3 Count the number of surviving juveniles every day for 6 days. The survival rate curve was drawn by GraphPad Prism software.

[0058] Experi...

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Abstract

The invention discloses nucleic acid molecule, an expression vector and an application thereof, an antibacterial drug and a method thereof, which belong to the technical field of biology. A nucleotidesequence of the nucleic acid molecule disclosed in the present invention is shown as SEQ ID NO. 1. The study of the present invention finds that the nucleic acid molecule shown in SEQ ID NO. 1 is overexpressed in fish body, which can significantly inhibit bacterial proliferation and increase the survival rate of the fish body during bacterial infection; for example, the nucleic acid molecule shown as SEQ ID NO. 1 is overexpressed in zebrafish, which can significantly inhibit the proliferation of edwardsiella tarda in zebrafish and significantly improve the survival rate of zebrafish during infection with edwardsiella tarda; the nucleic acid molecule shown as SEQ ID NO. 1 has broad application prospects in the field of aquaculture, can be used for preparing antibacterial drugs or disease-resistant biological products, and improves the antibacterial ability of fish body.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a nucleic acid molecule, an expression vector and its application, an antibacterial drug and a method. Background technique [0002] Histones are important components of chromosome nucleosomes; according to their molecular weight and amino acid composition, histones can be divided into H1, H2A, H2B, H3 and H4. Histone H1 is particularly rich in lysine; H2A and H2B are rich in lysine; H3 and H4 are rich in arginine. Among the five histones, the 8-mer composed of two molecules each of H2A, H2B, H3 and H4 is the core part of the nucleosome; while H1 is involved in binding DNA between nucleosomes and forming higher-order structures. Histones are the most conserved of known proteins. In species with distant relatives, histone H1 changes greatly, and H3 and H4 change the least; for example, there are only 3 amino acid differences between the amino acid sequence of higher vertebrate human...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/46C12N15/85A01K67/027A61K38/17A61P31/04
CPCA01K67/0276A01K2217/00A01K2227/40A01K2267/02A61K38/1706A61P31/04C07K14/461C12N15/85
Inventor 昌鸣先武小曼曹璐
Owner INST OF AQUATIC LIFE ACAD SINICA
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