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Colibacillus gene engineering bacterin K88-K99-LT for firstborn piglet diarrhea and construction method thereof

A K88-K99-LT, genetic engineering vaccine technology, applied in genetic engineering, biochemical equipment and methods, plant genetic improvement, etc.

Inactive Publication Date: 2008-02-13
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At home and abroad, there is no genetically engineered vaccine product that expresses three of the above six protective antigen genes in tandem

Method used

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  • Colibacillus gene engineering bacterin K88-K99-LT for firstborn piglet diarrhea and construction method thereof
  • Colibacillus gene engineering bacterin K88-K99-LT for firstborn piglet diarrhea and construction method thereof

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Experimental program
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Effect test

Embodiment Construction

[0031] 1. Biomaterials:

[0032] 1. Protective antigen sequence

[0033] The gene sequences of the main subunits of the above three protective antigens are from GenBank of the Internet, among which:

[0034] ①The accession number of faeG(K88) in GenBank is M29375, and the accession time is 1993-4-26.

[0035] ② The accession number of fanC(K99) in GenBank is M35282, and the accession time is 1993-4-26.

[0036] ③The accession number of ltb(LT) in GenBank is M17873, and the accession time is 1996-4-16.

[0037] 2. Expression plasmid

[0038] The expression plasmid is pGEX-6P-1, purchased from Amersham Pharmacia.

[0039] 3. The source of the strain

[0040] The recombinant plasmid containing the protective antigen gene is expressed in BL21 host bacteria. BL21 host bacteria is also a commercial host bacteria, also purchased from Amersham Pharmacia.

[0041] 2. Construction process:

[0042] 1. Obtaining protective purpose fragments

[0043] According to the gene sequenc...

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Abstract

The invention relates to Escherichia coli genetic engineering vaccine 987P-ST-F41 for curing diarrhea of baby pigs and the constitution method, pertaining to the field of veterinary technology. The major subunit gene faeG segment of the expanded K88, the major subunit gene fanC segment of K99 and the major subunit gene ltb segment of LT linked to the pronucleus expression vector pGEX-6P-1 in series in a faeG-fanC-ltb manner; after transformation of host bacteria BL21, IPTG induction expression is conducted, and proved by SDS-PAGE and Western-blot that the objective gene is expressed. The vaccine is capable to prevent diarrhea of baby pigs, and the safety and immunogenicity are reliable.

Description

technical field [0001] The invention relates to the development of an Escherichia coli vaccine for neonatal piglet diarrhea, and belongs to the technical field of animal husbandry and veterinary medicine. Background technique [0002] Neonatal diarrhea caused by a specific serotype of Enterotoxigenic Escherichia coli (ETEC) is an important infectious disease that leads to the death of newborn piglets. Diarrhea in newborn piglets often manifests as sudden onset, yellow watery feces, rapid dehydration and electrolyte imbalance. Affected pigs can die within a few hours, and some piglets have died without diarrhea. Escherichia coli of a specific serotype is the main pathogen that causes diarrhea in newborn piglets, and it mainly produces four types of adhesins, F4 (K88), F5 (K99), F6 (987P) and F41, and can also produce heat-stable enterotoxins (heat-stable enterotoxins). enterotoxin, ST) or / and heat-labile enterotoxin (LT). In view of the high occurrence rate of four types o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/33A61K39/108A61P31/04
Inventor 高崧刘秀梵马林艳
Owner YANGZHOU UNIV
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