Mermaid luminous bacillus outer membrane protein V and coding sequence, its preparation method and application

A technology of photobacteria and outer membrane protein, applied in the field of preparation of polynucleotides and polypeptides

Inactive Publication Date: 2007-10-24
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has not been reported that the Bacillus OmpV protein has a salt-sensitive function

Method used

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  • Mermaid luminous bacillus outer membrane protein V and coding sequence, its preparation method and application
  • Mermaid luminous bacillus outer membrane protein V and coding sequence, its preparation method and application
  • Mermaid luminous bacillus outer membrane protein V and coding sequence, its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Cloning of Embodiment 1 Photobacterium mermaid OmpV gene

[0070] 1. Acquisition and cultivation of bacteria

[0071] Various bacteria obtained from sea areas, after simple separation and purification, were initially screened with Vibrio special medium TCBS to obtain a green colony, which was identified as mermaid luminescence after identification by VITEK32 automatic bacteria identification instrument of Xiamen Center for Disease Control and Prevention bacilli.

[0072] Pick a single colony of Photobacterium mermaidi and culture it overnight in LB medium, then add 30% glycerol and store it at -70°C.

[0073] 2. Cloning of full-length DNA

[0074] Since the complete sequence of the gene of Photobacter mermaidus has not been determined, there is no report of the gene sequence of OmpV of this bacterium at present. In view of the complete sequence determination of Vibrio parahaemolyticus, we designed OmpV gene primers according to its OmpV sequence for PCR amplification...

Embodiment 2

[0092] Example 2 Sequence information and homology analysis of Photobacterium mermaid OmpV gene

[0093] The full length of the novel Photobacterium mermaid OmpV gene of the present invention is 777bp, and the detailed sequence is shown in SEQ ID NO.3. According to the full length of the DNA, the amino acid sequence of Photobacterium mermaid OmpV was deduced, with a total of 258 amino acid residues and a molecular weight of 27kD. The detailed sequence is recorded as SEQ ID NO.4, and the relevant information is:

[0094] (i) Sequential features:

[0095] (A) Length: 258 amino acids

[0096] (B) Type: amino acid

[0097] (C) chain: single chain

[0098] (D) Topology: Linear

[0099] (ii).Molecular type: polypeptide

[0100] (iii). Sequence description:

[0101] MNKTLALLT LAAAGTATAG DTYIRNGNIY NNQGGWVAEV GVAQGSDLFK DQKHNTAPIL 60

[0102] NGGYHGEDFN ADLGGTNYRF LGETNDLINM SAYVVGSGII RDGDVAKSLK GTQKRRLAVD 120

[0103] LGLNTDFTLD EHNVISTYLQ HDISGAYKGY LAGATYFHIM NFGSVDFVPF ANLT...

Embodiment 3

[0171] Example 3 Research on the structure and function of Photobacterium mermaid OmpV protein

[0172] The changes of Photobacterium mermaid OmpV under different salt concentration conditions are given below:

[0173] After extracting the outer membrane proteins of Photobacterium mermaidi under the culture conditions of three salt concentrations of 0.5%, 1% and 4%, firstly, the outer membrane protein profiles of them were compared and analyzed by SDS-PAGE, and the results are shown in Figure 1 . It can be clearly seen from Figure 1 that with the gradual increase of salinity, a protein band also shows a gradual decrease in the amount, which indicates that this protein band is compatible with the salt adaptability of Photobacterium mermaidi are closely related. Combined with the identification results of the outer membrane protein of Photobacterium mermaidus, it can be known that this band is OmpV.

[0174] Cloning of Photobacterium mermaid OmpV protein and its effect on the...

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Abstract

The present invention provides a new mermaid photobacillus outer membrane protein V, nucleotide sequence coding polypeptide with protein activity of mermaid photobacillus outer membrane protein V, amino acid sequence of mermaid photobacillus outer membrane protein V and method for preparing mermaid photobacillus outer membrane protein V by utitizing recombination technique. Said invention also provides the application of mermaid photobacillus Omp V protein and coding sequence.

Description

technical field [0001] The present invention belongs to the field of biotechnology, and in particular relates to a new polynucleotide encoding outer membrane protein V (outer membrane protein V, OmpV) of Photobacterium damsela and the polypeptide encoded by the polynucleotide, and also relates to the Preparation methods and uses of polynucleotides and polypeptides. Background technique [0002] Bacillus is currently the most studied Gram-negative bacteria. Gram-negative bacteria have strong adaptability and stress resistance due to their unique cell wall structure. Gram-negative bacteria are mostly opportunistic pathogenic bacteria, which can cause a variety of infections when the body's immunity is low. Many diseases in humans and livestock are caused by Gram-negative bacteria. The main pathogenic bacilli include Shigella, Salmonella typhi, Escherichia coli, Proteus, Pseudomonas aeruginosa, Bacillus pertussis, etc. They produce endotoxins and make people sick through endot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/28C12N15/31C12P21/02C12N1/21
Inventor 彭宣宪吴丽娜
Owner SUN YAT SEN UNIV
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