Prokaryotic expression and phylogenetic analysis of ORF2 gene of goose astrovirus

doi:10.3969/j.issn.1004-1524.2020.05.05

›› 2020, Vol. 32 ›› Issue (5): 789-797.DOI: 10.3969/j.issn.1004-1524.2020.05.05

• Animal Science • Previous Articles Next Articles

Prokaryotic expression and phylogenetic analysis of ORF2 gene of goose astrovirus

PU Lusha^1,2, SU Shibo², CHEN Xiaohan², ZHAO Lili^2,*, CHEN Hongyan^2,*

1.College of Animal Science, Heilongjiang Bayi Agricultural University, Daqing 163000, China;
2. State Key Laboratory of Veterinary Biotechnology, Heilongjiang Provincial Key Laboratory of Laboratory Animal and Comparative Medicine, Harbin Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Harbin 150069, China

Received:2019-09-20 Online:2020-05-25 Published:2020-05-29

Abstract

Abstract: The goose astrovirus (GoAstV) ORF2 gene was amplified by RT-PCR, and the amplified product was cloned into the prokaryotic expression vector pET32a expressed in E.coli BL21. SDS-PAGE analysis showed that the ORF2 protein with the size of 106 ku was obtained after the induction of recombinant bacteria, which was consistent with the theoretical value. The polyclonal antibody was prepared by immunizing mice with the induced protein that was purified by Ni column affinity chromatography and the antibody titer was determined. Western blotting results showed that ORF2 protein could be recognized by mouse anti GoAstv positive serum. Sequence analysis showed that the nucleotide similarity between the virus and other astrovirus of goose origin were between 37.8% and 99.3%. The HLJ-1801 isolate was located in the same branch with the eight published strains of astrovirus of goose origin, indicating that it belonged to a new type of astrovirus. In this study, the GoAstV ORF2 protein was successfully and efficiently expressed using a prokaryotic expression system, and the fusion protein existed as an inclusion body. The purified ORF2 recombinant protein and mouse anti goastv polyclonal antibody were successfully obtained, which laid a foundation for further research on the diagnostic reagent of ORF2 protein for astrovirus infection.

Key words: goose astrovirus, ORF2, prokaryotic expression, Phylogenetic analysis

CLC Number:

S855.3

PU Lusha, SU Shibo, CHEN Xiaohan, ZHAO Lili, CHEN Hongyan. Prokaryotic expression and phylogenetic analysis of ORF2 gene of goose astrovirus[J]. , 2020, 32(5): 789-797.

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Prokaryotic expression and phylogenetic analysis of ORF2 gene of goose astrovirus

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