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
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[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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