Recombinant fusion protein of 8 proteins of streptococcus equi subsp.equi as well as preparation method and application of recombinant fusion protein

A technology of fusion protein and Streptococcus equi, which is applied in the field of recombinant fusion protein and its preparation, can solve problems such as adverse reactions, and achieve the effect of avoiding infection and high safety

Pending Publication Date: 2022-05-06
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Pinnacle IN attenuated vaccine is made from New York isolate CF32, and is prepared by nitrosoguanidine attenuation. The intramuscular injection of the vaccine will cause adverse reactions

Method used

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  • Recombinant fusion protein of 8 proteins of streptococcus equi subsp.equi as well as preparation method and application of recombinant fusion protein
  • Recombinant fusion protein of 8 proteins of streptococcus equi subsp.equi as well as preparation method and application of recombinant fusion protein
  • Recombinant fusion protein of 8 proteins of streptococcus equi subsp.equi as well as preparation method and application of recombinant fusion protein

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1 Expression, purification and immune efficacy experiment of recombinant fusion protein

[0040] Material

[0041] Streptococcus equi subspecies equi (S.equi) HLJ2018D-LX strain was isolated and preserved by the Equine Infectious Diseases and Lentivirus Research Innovation Team Laboratory of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences; E.coli BL21(DE3) , DH5α was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd. The pGex-6p-1 and pET-28a vectors are maintained by the laboratory.

[0042] method

[0043] 1. Amplification of the target gene

[0044] The nucleotide sequences of the primers are listed in Table 1.

[0045] Table 1

[0046]

[0047]

[0048]

[0049] Using the s.equi HLJ2018D-LX genome as a template, cneSEco(6p-1)F-cneSXho(6p-1)R, eagSEco(6p-1)F-eagSXho(6p-1)R, SclFsEco(6p-1)R, respectively )F-SclFsXho(6p-1)R, SclIsEco(6p-1)F-SclIsXho(6p-1)R, SclCsEco(6p-1)F-SclCsXho(6p-1)R, eq8sEco(...

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Abstract

The invention discloses a recombinant fusion protein of 8 proteins of streptococcus equi subsp.equi as well as a preparation method and application of the recombinant fusion protein. The recombinant fusion protein is obtained by sequentially connecting eight proteins of cne, eag, SclC, SclI, SclF, eq5, eq8 and ideE of streptococcus equi subsp.equi through protein connectors Gly-Gly-Gly. Preferably, the recombinant fusion protein is named as SE8, and the amino acid sequence of the recombinant fusion protein is as shown in SEQ ID NO. 2. Experiments prove that the recombinant fusion protein SE8 prepared by the invention has good antigenicity and immunogenicity, an immunized mouse can effectively induce an organism to generate a high-level specific antibody, and the recombinant fusion protein SE8 plays a role in protecting an S.equi infected mouse. In addition, the recombinant fusion protein is a mycoprotein prepared by purifying escherichia coli, does not contain other components, has high safety, and avoids the phenomenon of infection.

Description

technical field [0001] The present invention relates to a recombinant fusion protein and its preparation method and application, in particular to a recombinant fusion protein composed of 8 proteins of Streptococcus equi subsp. Applications. The invention belongs to the technical field of veterinary medicine. Background technique [0002] Equine blight is a common infectious disease of horses and donkeys. It is a three-category animal disease stipulated in my country. At present, cell-free vaccines produced by many foreign companies are being put into use, including Strepvax II (produced by Boehringer Ingelheim, Germany), Equivac S (produced by New Zealand Zoetis), Strepguard (produced by MSDAnimalHealth), etc. These vaccines are administered intramuscularly, but the effectiveness of these vaccines is currently uncertain. [0003] seM protein is an important virulence protein of Streptococcus equi subsp. Association[J], 235.), so many vaccine studies start with this prote...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70C12N15/66C12P21/02A61K39/09A61P31/04
CPCC07K14/315C12N15/70A61K39/092A61P31/04C07K2319/00A61K2039/552
Inventor 郭巍王晓钧杜承胡哲
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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