A kind of method for preparing tetramer of O-type foot-and-mouth disease virus polypeptide

Abstract

The invention belongs to the field of biotechnology, and in particular relates to a method for preparing a tetramer of an O-type foot-and-mouth disease virus polypeptide. For the designed foot-and-mouth disease virus polypeptide, the present invention will adopt two methods of ELISPOT and construction of tetramer to screen and identify the foot-and-mouth disease virus polypeptide, construct the tetramer of the heavy chain of purse pig SLA-2, and use it in the prokaryotic expression system pET-21a(+) / BL21 was expressed and inclusion bodies were extracted; foot-and-mouth disease virus peptides were screened for peptides that could cause specific CTL reactions by ELISPOT method, and then the peptides were refolded and biotinylated with the heavy chain of SLA‑2 and the existing light chain β2m And react with FITC-labeled streptavidin to generate SLA-2-BSP / peptide tetramer, and then detect the function of tetramer by flow cytometry. The present invention obtains a specific polypeptide epitope of porcine O-type foot-and-mouth disease virus VP1 through the ELISPOT method identification, and successfully constructs a peptide tetramer, and preliminarily establishes a porcine tetramer technology platform for the study of porcine-specific CTL immune response and T epitope screening lays the foundation.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing a tetramer of an O-type foot-and-mouth disease virus polypeptide. Background technique [0002] Foot-and-mouth disease is an acute febrile high-contact infectious disease caused by foot-and-mouth disease virus (FMDV), which mainly affects cloven-hoofed animals, and pigs are more susceptible groups. Blisters and festering are the main features, and it is the most contagious type of livestock disease. Every outbreak of foot-and-mouth disease will bring great losses to the agricultural economy. Therefore, the International Bureau of Animal Diseases (OIE) has always listed it as the top of the list of Category A animal diseases that must be reported. Therefore, the prevention and diagnosis of foot-and-mouth disease in pigs is particularly important. At present, the prevention and treatment of foot-and-mouth disease virus mainly relies on traditional i...

AI Technical Summary

Problems solved by technology

[0006] At present, the Tetramer technology has not been applied to the identification of the function of the foot-and-mouth disease virus polypeptide bound by the porcine SLA class I molecule in China.

Images