Swine trichinella spiralis vaccine as well as preparation method and application thereof

A technology for swine trichinosis and trichinella, applied in the biological field, can solve the problems that ES antigens are difficult to ensure stability, difficult to distinguish effective antigen components or purification treatment, etc., and achieve the effect of good immune protection and great application prospects.

Inactive Publication Date: 2018-01-09
SUN YAT SEN UNIV
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Problems solved by technology

Since Trichinella spiralis is still unable to be subcultured in vitro, it is difficult to ensure the stability of the obtained ES antigen, and the ES antigen is a mixture of various antigens, so it is difficult to distinguish the effective antigenic components or purification treatment

Method used

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  • Swine trichinella spiralis vaccine as well as preparation method and application thereof
  • Swine trichinella spiralis vaccine as well as preparation method and application thereof
  • Swine trichinella spiralis vaccine as well as preparation method and application thereof

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Embodiment 1

[0046] Embodiment 1 A kind of preparation method of trichinella spiralis vaccine

[0047] 1, TsCPB2 gene cloning

[0048] Total RNA was extracted from muscle larvae 35 days after infection of mice using TRIZOL (Invitrogen, USA) reagent. Total RNA was reverse-transcribed into cDNA first-strand using reverse transcriptase (M-MLV RT, Promega). by TsCPB2 EST sequence (AccessionNo. EX501780) designed gene-specific amplification primers, TsCPB2-5’GSP outer primer: 5’-CGGATCTGTTGTCCTGGTTGG-3’; TsCPB2-5’GSP inner primer: 5’-CTGTTGTCCTGGTTGGATAAA-3’. The full sequence of TsCPB2 was amplified by 3'-RACE-PCR (TaKaRa, Japan), and the operation was performed according to the method recommended by the kit. PCR products were electrophoresed on 1% agarose gel, and PCR products were recovered using QIAquick gel extraction kit (QIAGEN, USA). The PCR product was cloned into the pMD19-T vector and sequenced to obtain TsCPB2 Nucleotide sequence ( figure 1 ), its sequence is shown in SE...

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Abstract

The invention discloses a swine trichinella spiralis vaccine as well as a preparation method and an application thereof. Firstly, a mature region of a gene TsCPB2 is amplified and cloned into an expression vector of a His tag, and then after transformation of escherichia coli and IPTG induced expression, TsCPB2 recombinant protein is purified by Ni-affinity chromatography, and a trichinella spiralis excretion-secretion recombinant antigen is obtained; TsCPB2 recombinant protein and an immune adjuvant are subjected to mixed emulsification, and the TsCPB2 recombinant antigen vaccine is obtained.With adoption of the vaccine for immunizing a mouse or swine, the host can be induced to produce an immune reaction, a good immunoprotective effect on trichinella spiralis infection is realized, andgreater application prospect is achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, it relates to a porcine trichinosis vaccine and its preparation method and application. Background technique [0002] Trichinella ( Trichinella spiralis ) is a zoonotic parasitic nematode that can parasitize more than 150 mammals including humans, pigs, dogs, mice, and cats, causing trichinosis. The mutual infection of pigs and rats is an important source of human trichinellosis, and pigs are the main source of animal infection. Raw or half-cooked pork infected with Trichinella spiralis larva cysts is the main way for people to be infected with Trichinella spiralis. Trichinellosis is distributed worldwide, mainly manifested as fever, edema and other symptoms, followed by symptoms such as muscle pain and limb weakness, and severe cases can lead to death. In recent years, epidemics and outbreaks of the disease have occurred in many parts of the world. In my country, Yunnan, Hena...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61P33/10C12N15/70C12N15/57C12N9/64C07K16/40C07K16/18
Inventor 廖婉琴周兴旺
Owner SUN YAT SEN UNIV
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