禽呼肠孤病毒σB和σC蛋白在昆虫细胞中的共表达

doi:10.16431/j.cnki.1671-7236.2020.05.006

Abstract

Abstract: To obtain the native conformation and antigenicity of σB and σC proteins of avian reovirus,the σB and σC proteins was expressed by Bac-to-Bac baculovirus expression system,and its reactivity was identified.The σB and σC genes of avian reovirus were amplified by designing two pairs of primers according to the sequence published on NCBI,inserted into donor plasmid pFast-dual to obtain recombinant donor plasmid,then transformed into E.coli DH10Bac,and the recombinant bacmid plasmid was obtained.Recombinant baculovirus was transfected into sf9 insect cells using cellfection reagent to obtain the production of recombinant viral contained the sequence of σB and σC genes.sf9 insect cells were inoculated by recombinant viral to explore the MOI and harvest timing expression condition,and the expression condition was optimized.The results of Western blotting and indirect immunofluorescence assay (IFA) showed that the recombinant σB and σC proteins were expressed successfully in sf9 insect cells and had good biological activity.This study provided a further research direction of ARV vaccine of virus-like particles and ARV subtype antibody ELISA detection kit.

Key words: avian reovirus (ARV); σB gene; σC gene; baculoviruses expression system; sf9 cell

CLC Number:

S831.7

WANG Sheng, XIE Zhixun, SHEN Wenkang, XIE Liji, FAN Qing, LUO Sisi, ZHANG Yanfang, ZENG Tingting, HUANG Jiaoling, ZHANG Mingxiu, XIE Zhiqin, DENG Xianwen. Expression of σB and σC Proteins of Avian Reovirus in Insect Cells[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(5): 1334-1341.

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Expression of σB and σC Proteins of Avian Reovirus in Insect Cells

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