Attenuated vector bacterium of salmonella choleraesuls and construction method of attenuated vector bacterium

A Salmonella, construction method technology, applied in the field of animal bacterial genetic engineering, to achieve the effects of strong immunogenicity, high safety, and improved immune efficiency

Inactive Publication Date: 2017-05-31
YANGZHOU UNIV
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Problems solved by technology

Deleted in Salmonella choleraesuis sopB Whether the gene will achieve the same effect of reducing the inflammatory response caused by Salmonella choleraesuis infection and improving the immunogenicity of Salmonella choleraesuis to pigs has not been reported so far at home and abroad

Method used

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  • Attenuated vector bacterium of salmonella choleraesuls and construction method of attenuated vector bacterium
  • Attenuated vector bacterium of salmonella choleraesuls and construction method of attenuated vector bacterium
  • Attenuated vector bacterium of salmonella choleraesuls and construction method of attenuated vector bacterium

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Embodiment Construction

[0081] In order to clarify the technical scheme and technical purpose of the present invention, the present invention will be further introduced below in conjunction with the drawings and specific embodiments.

[0082] 1. Construction of an attenuated vector of Salmonella choleraesuis:

[0083] 1. Construct a sopB Suicide vector and identification of mutant gene:

[0084] According to the complete gene sequence of Salmonella cholerae suis in Genbank, find the Δ sopB The entire gene sequence of the gene and its upstream and downstream sequences, primers P1, P2, P3, and P4 are designed according to the upstream and downstream genes of the gene, and the 300bp or so fragments of the upstream and downstream genes are homology arms, knock out sopB Gene (ATG to TAA), such as figure 1 Shown.

[0085] The specific steps are as follows: Using the genome of Salmonella choleraesuis C78-3 as a template, it was amplified by PCR with primers P1 and P2 pipC The homologous arm gene sequence, calle...

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Abstract

The invention provides an attenuated vector bacterium of salmonella choleraesuls and a construction method of the attenuated vector bacterium and belongs to the technical field of animal bacterium genetic engineering. The attenuated vector bacterium is C78-3 salmonella choleraesuls without deltamanA, deltacrp::TT araC PBAD, deltarelA:: araC PBAD lacI TT, deltasoB and deltaasdA genes and is named as rSC0016. The method for taking a suicide vector without a resistance marker as a salmonella choleraesuls constructing vector is adopted and the utilization of the vector marked with antibiotics can be avoided; the vector can be safely used in clinical practice; furthermore, a balanced lethal system carries an exogenous antigen and can guarantee that only vaccine strains carrying the exogenous antigen can be used for producing vaccines and enter a host and survive, so that the immunity efficiency of the vaccines is improved.

Description

Technical field [0001] The invention belongs to the technical field of animal bacterial genetic engineering. Background technique [0002] Salmonella typhimurium weakened by genetic engineering methods is used as a vector to express foreign genes and is widely used in the treatment of various human and animal diseases, and has achieved good results. However, the Salmonella vectors in many reports either overly attenuate the attenuated vectors and lose their immunogenicity, or although they retain good immunogenicity, the carrier strains are not attenuated enough and lack safety. [0003] Salmonella choleraesuis is an important pathogen of pigs. There have been a few reports on the delivery of other pathogenic antigens of pigs with Salmonella choleraesuis as a carrier. For example, the laboratory of Academician Chen Huanchun of Huazhong Agricultural University uses Salmonella choleraesuis Chinese attenuated vaccine standard strain C500 as a carrier to deliver Mycoplasma hyopneumoni...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/74C12R1/42
Inventor 石火英李玉安吉贞颖尚竞吕一帆
Owner YANGZHOU UNIV
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