V. alginolyticus outer-membrane protein W gene, process for preparing the same and uses thereof

A kind of technology of Vibrio alginate and Alginic acid arc, which is applied in the field of genes and can solve the problems such as salt tolerance function of Vibrio OmpW protein that have not been reported.

Inactive Publication Date: 2005-11-23
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It has not been reported that Vibri...

Method used

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  • V. alginolyticus outer-membrane protein W gene, process for preparing the same and uses thereof
  • V. alginolyticus outer-membrane protein W gene, process for preparing the same and uses thereof
  • V. alginolyticus outer-membrane protein W gene, process for preparing the same and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Cloning of Vibrio alginolyticus OmpW gene

[0060] 1. Bacterial culture

[0061] Vibrio alginolyticus was derived from the strains preserved in our laboratory, and a single colony was picked and cultured overnight in LB medium, then stored at -70°C with 30% glycerol.

[0062] 2. Cloning of full-length DNA

[0063] The Vibrio alginolyticus OmpW can be successfully identified by using the genome data of Vibrio parahaemolyticus as the search object, but from the point of view that the significance of the results is not high, there should be a sequence difference between the two bacteria. The differences but similarities are still high. Thus, primers can be designed according to the 5' end and 3' end of the Vibrio parahaemolyticus OmpW sequence, plus a restriction site and two protective bases, R1: 5'-GGGGATCCATGAAAAAAACAATCTG-3' (denoted as SEQ ID NO .1) is the forward primer, oligonucleotide R2: 5'-CCGAATTCTTAGAACTTGTAACCGC-3' (denoted as SEQ ID NO.2) is the ...

Embodiment 2

[0081] Example 2 Sequence information and homology analysis of the OmpW gene of Vibrio alginolyticus

[0082] The full length of the new OmpW gene of vibrio alginilyticus of the present invention is 642 bp, and the detailed sequence is shown in SEQ ID NO.3. The amino acid sequence of Vibrio alginolyticus OmpW was deduced based on the full length of the DNA, with a total of 213 amino acid residues and a molecular weight of 22Kd. The detailed sequence is recorded as SEQ ID NO.4, and the relevant information is:

[0083] (i) Sequential features:

[0084] (A) Length: 213 amino acids

[0085] (B) Type: amino acid

[0086] (C) chain: single chain

[0087] (D) Topology: linear

[0088] (ii).Molecular type: polypeptide

[0089] (iii). Sequence description:

[0090] 1 MKKTICSLAV VAALVSPSVF AHKQGDFVLR VGAASVVPND SSDKILGSQE ELEVDSNTQL

[0091] 61 GLTFGYMFTD NISLELLAAT PFSHDISTDL VGSDIAKTKH LPPTLMVQYY FGESQSKFRP

[0092] 121 YVGAGLNYTI FFDEDFNSTG KGADLSDLKL DDSFGLAANI GVDYMINDQW F...

Embodiment 3

[0150] Example 3 Research on the structure and function of the OmpW protein of Vibrio alginolyticus

[0151] The change of Vibrio alginolyticus OmpW under different salt concentration conditions is given below:

[0152] After extracting the outer membrane proteins of Vibrio alginolyticus under four kinds of salt concentration culture conditions of 1%, 2%, 3% and 4%, firstly, compare and analyze their outer membrane protein profiles by SDS-PAGE, the results are as follows figure 1 shown. From figure 1 It can be clearly seen that with the gradual increase of salinity, there is a protein band that also shows a gradual increase in the amount, which indicates that this protein band is closely related to the salt adaptation of Vibrio alginolyticus sex. Combined with the identification results of the outer membrane protein of Vibrio alginolyticus, it can be known that this band is OmpW.

[0153] The change of Vibrio parahaemolyticus OmpW under different salt concentration condi...

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Abstract

The invention relates to a V. alginolyticus outer-membrane protein W gene, process for preparing the same and uses. The invention provides a novel V.alginolyticus OmpW gene, and the use of V.alginolyticus and Vibrio vulnficus OmpW protein and coded sequence, which comprises, linking purified nucleic acid sequence encoding protein active polypeptides having V.alginolyticus OmpW to expression regulation sequence, forming V.alginolyticus OmpW protein expression carrier, transferring the expression carrier into host cells, forming the recombination cells of the V.alginolyticus Omp protein, culturing the recombination cells and separating.

Description

technical field [0001] The invention relates to a gene, in particular to an outer membrane protein W (outer membrane protein W, OmpW) gene expressed on the outer membrane of Vibrio alginolocius, a preparation method and use thereof. Background technique [0002] Vibrio is an important class of marine pathogenic bacteria. It is the main pathogen of gastrointestinal diseases in coastal areas. At the same time, they are also important pathogens in aquaculture. The occurrence of vibriosis often leads to huge economic losses. loss. The ability to adapt to different salinity environments is a basic function of marine pathogenic microorganisms. At the same time, it is also closely related to the pathogenicity and reproduction ability of bacteria. The outer membrane protein is located in the outermost layer of bacteria and directly communicates with the external environment. (including both the marine environment and the host's internal environment), and in-depth research on it is ...

Claims

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Application Information

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IPC IPC(8): C12N15/31
Inventor 彭宣宪吴丽娜徐常新王三英
Owner XIAMEN UNIV
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