山羊痘病毒毒力因子KLP2的表达及多克隆抗体的制备

doi:10.16431/j.cnki.1671-7236.2017.10.004

Abstract

Abstract:

In order to study capripox virus, and prepare polyclonal antibody of its virulence factor KLP2, the prokaryotic expression vector of two structure domains (KLP2-1 and KLP2-2) were built with the method of homologous recombination genes and identified by sequencing. The plasmid were transformed into E. coli BL21 and inducing expressed by IPTG, objective protein were tested by SDS-PAGE and Western blotting, and purified by KCl dyeing rubber cutting.The rabbits were immuned with freund's adjuvant, the prepared antibody were evaluated by Western blotting and ELISA test. The results showed that the expression vector of pET42b-KLP2-1 and pET42b-KLP2-2 were built successfully, and KLP2-1 and KLP2-2 genes were 657 and 984 bp, and expressed about 27 and 38 ku protein, respectively, it was consistent with theories value. The concentrations of rubber cutting purified protein was 1 to 2 mg/mL. The antibody showed obvious specific bands detecting by Western blotting, and its titer were 1:512 detecting by ELISA. This result indicated that KLP2-1 and KLP2-2 proteins were expressed successfully, and the corresponding polyclonal antibody were prepared.

Key words: capripox virus; KLP2 gene; protein expression; antibody preparation

CLC Number:

S852.65

LI Jin-na, SONG Shu-ting, CHEN Hong-wei, LIU Tao, ZHAO Ai-yun, LI You-wen. Expression of Goatpox Virus Virulence Factor KLP2 and Preparation of Polyclonal Antibody[J]. , 2017, 44(10): 2851-2857.

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Expression of Goatpox Virus Virulence Factor KLP2 and Preparation of Polyclonal Antibody

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