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Polypeptide and DNA combined vaccine for resisting animal's foot-and-mouth disease and its preparing method

A technology of DNA vaccine and polypeptide vaccine, which is applied in the field of genetic engineering, can solve the problems of slow immune response, low antigen expression and weak antigen expression

Inactive Publication Date: 2003-04-09
FUDAN UNIV
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

The disadvantage of DNA vaccines is that the antigen expression is not high, and the immune response is relatively slow and weak

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0027] The present invention will be further specifically described below by taking the combination vaccine of porcine foot-and-mouth disease O-type polypeptide and DNA as an example.

[0028] The construction process of peptide vaccine. The DNA sequence encoding the 200ヘ213 and 141ヘ160 amino acid fragments in the VP1 gene and the linker DNA sequence were synthesized by chemical methods, and concatenated at the gene level to form a primary structure of 141ヘ160AA-200ヘ213AA-141ヘ160AA (SEQ ID NO.1), and insert it into the cloning vector pBluescriptIIKS. Take 1 ug of pBluescriptIIKS and digest it with EcoR I and BamH I, react at 37°C for two hours, and recover large fragments by electrophoresis and gel slicing. The large fragment DNA is mixed with the above-mentioned tandem fragments, and a ligation reaction occurs under the action of T4 ligase and the like. The obtained recombinant DNA was transformed into Escherichia coli competent cells, cultured on ampicillin plates, and inv...

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Abstract

The combined vaccine consists of a kind of polypeptide vaccine and one DNA vaccine, and may be used to immunize suscaptible animal in several modes to prevent foot-mouth disease virus infection. The chemically synthetic encoding foot-mouth disease virus VP1 protein has the DNA sequence of antigen amino acid peptide segment and is connected into multicopy serial structure. The antigen gene with serial structure is then connected to beta-galactosidase gene to form fusion gene. When the fusion gene is inserted into protokaryon expression vector, the expressed fusion protein may be compounded into the polypeptide vaccine. When the fusion gene is inserted into eukaryon expression vector, the recombinant eukaryon expression plasmid may be amplified to obtain NDA vaccine.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a combined vaccine of polypeptide and DNA against domestic animal foot-and-mouth disease and a preparation method thereof. Background technique [0002] Foot-and-mouth disease (FMD) is one of the most serious livestock infectious diseases in the world today, mainly harming pigs, cattle, sheep and other cloven-hoofed animals, and has a wide range of outbreaks, rapid spread, and high mortality after infection , causing huge economic losses to the animal husbandry industry, and thus was listed as the first class A infectious disease by the International Veterinary Bureau. [0003] Genetic engineering polypeptide vaccines have broad application prospects due to their advantages of good safety, easy storage and ideal effect. However, secondary immunization is necessary to better stimulate the body's immune system so that it has sufficient resistance to foot-and-mouth di...

Claims

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Application Information

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IPC IPC(8): A61K39/135A61K48/00A61P31/12C12N15/63
Inventor 郑兆鑫严维耀李光金陈维灶刘明秋
Owner FUDAN UNIV
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