Prokaryotic expression of recombinant early pregnancy factor and preparation of polyclonal antibody

doi:10.14088/j.cnki.issn0439-8114.2023.05.023

HUBEI AGRICULTURAL SCIENCES ›› 2023, Vol. 62 ›› Issue (5): 129-134.doi: 10.14088/j.cnki.issn0439-8114.2023.05.023

• Animal Husbandry & Veterinary Medicine • Previous Articles Next Articles

Prokaryotic expression of recombinant early pregnancy factor and preparation of polyclonal antibody

WANG Cong¹, WANG Han¹, ZHANG Hai-ling², XIAO Yi-ran¹, GUO Yu-xi¹, LIU Meng-di¹, LI Hao-song¹, WU Zong-cheng¹, DUAN Fu-chun¹, LU Shi-ying¹

1. College of Veterinary Medicine/Key Laboratory of Zoonosis Research,Ministry of Education,Jilin University,Changchun 130112,China;
2. Institute of Specialty, Chinese Academy of Agricultural Sciences,Changchun 130112,China

Received:2023-02-16 Online:2023-05-25 Published:2023-06-12

Abstract

Abstract: Bioinformatics analysis of bovine early pregnancy factor was conducted, and prokaryotic expression vector pET-28a-srEPF containing pro-soluble labels was constructed. The prokaryotic expression vector was successfully induced by IPTG, and the early pregnancy factor was purified by affinity chromatography. Then, recombinant protein was used as immunogen to immunized animals to prepare polyclonal antibodies. The expressed proteins and their polyantibodies were detected by Western blotting. The results showed that the recombinant protein of early pregnancy factor had been successfully prepared, and its molecular weight was about 26 ku. After purification, the concentration was determined by BCA method to be 1.1 mg/mL. Both the recombinant protein of early pregnancy factor and EPF protein after enzyme digestion could be specifically bound to the prepared polyclonal antibody, and the titer determined by indirect ELISA was 1:256 000.

Key words: early pregnancy factor, prokaryotic expression, polyclonal antibodies

CLC Number:

WANG Cong, WANG Han, ZHANG Hai-ling, XIAO Yi-ran, GUO Yu-xi, LIU Meng-di, LI Hao-song, WU Zong-cheng, DUAN Fu-chun, LU Shi-ying. Prokaryotic expression of recombinant early pregnancy factor and preparation of polyclonal antibody[J]. HUBEI AGRICULTURAL SCIENCES, 2023, 62(5): 129-134.

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Prokaryotic expression of recombinant early pregnancy factor and preparation of polyclonal antibody

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