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Streptococcus equi subsp. Zooepidemicus gene deletion strain as well as preparation method and application thereof

A gene deletion, Streptococcus equi technology, applied in the field of genetic engineering, can solve the problem of low immunogenicity

Pending Publication Date: 2021-06-22
FOSHAN UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Subunit vaccines refer to vaccines made of certain surface structural components (antigens) of microorganisms that do not contain viral nucleic acids, but can induce the body to produce antibodies. Its components are only a few main surface proteins, which can avoid the production of many irrelevant antigens The antibodies induced by the vaccine can reduce the side effects of the vaccine and the related diseases caused by the vaccine, but in general, the immunogenicity of subunit vaccines is low, and other means are needed to improve the immune effect

Method used

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  • Streptococcus equi subsp. Zooepidemicus gene deletion strain as well as preparation method and application thereof
  • Streptococcus equi subsp. Zooepidemicus gene deletion strain as well as preparation method and application thereof
  • Streptococcus equi subsp. Zooepidemicus gene deletion strain as well as preparation method and application thereof

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Embodiment 1

[0036] 1. Primer design and synthesis

[0037] According to the whole genome sequence of Streptococcus equi subsp. zooepidemicus ATCC 35246 registered in GenBank (NO.CP002904.1), using the genomic DNA of Streptococcus equi subsp. Arm primers were synthesized by Shanghai Sangon Biotechnology Co., Ltd. See Table 1 for primer information.

[0038] Table 1 Primers and sequence information

[0039]

[0040] 2. Homologous recombination plasmid pG + Construction of host5-pgm-LR

[0041]The whole genome DNA of SEZ C55138 was extracted as a PCR template, and primers pgmL-f, pgmL-r were used to amplify the upstream homology arm of the pgm gene (pgmL) and primers pgmR-f, pgmR-r to amplify the downstream homology arm of the pgm gene (pgmR) , The upper and lower homology arm fragments were respectively connected with 19T-Vector and transformed to obtain recombinant plasmids 19T-pgm-L and 19T-pgm-R.

[0042] Digest 19T-pgm-L plasmid and pG with restriction endonuclease + host5 plas...

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Abstract

The invention relates to the technical field of gene engineering, in particular to a streptococcus equi subsp. Zooepidemicus gene deletion strain as well as a preparation method and the application thereof. The gene deletion strain is a pgm gene deleted streptococcus equi subsp. Zooepidemicus strain. A gene-deleted strain delta pgm is obtained by knocking out a pgm gene in a streptococcus equi subsp. Zooepidemicus strain through a homologous recombination technology. The gene deletion strain can effectively activate Caspase-1 and reduce the expression level of IL-18 so as to weaken the pathogenicity of the Caspase-1, has a good application prospect as a vaccine, and is of great significance in research of SEZ pathogenic mechanisms.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a Streptococcus equi subsp. zooepidemicus gene deletion strain and a preparation method and application thereof. Background technique [0002] Streptococcus equi ssp. zooepidemicus (Streptococccus equi ssp.zooepidemicus; SEZ) belongs to Group C Streptococcus and is an important pathogen of swine streptococcosis in my country, causing arthritis, meningitis, mastitis, sepsis, etc. in infected animals. caused huge economic losses. At present, vaccine immunization is the primary measure to effectively prevent and treat streptococcal suis. The clinical vaccines for streptococcal suis mainly include inactivated vaccines and subunit vaccines. [0003] Inactivated vaccines refer to culturing viruses or bacteria first, and then inactivating them by physical or chemical means. They can be composed of whole viruses or bacteria, or split vaccines can be composed of their split fr...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/74A61K39/09A61P31/04C12R1/46
CPCC07K14/315C12N15/746A61K39/092A61P31/04
Inventor 张熙李国潮付强马春全何颍欣林德锐杨傲冰陈坚涂玉蓉
Owner FOSHAN UNIVERSITY