牛轮状病毒UK株NSP1基因序列分析及原核表达

doi:10.16431/j.cnki.1671-7236.2015.05.011

Abstract

Abstract: Rotavirus is a major cause of acute diarrhea in both many kinds of young animals and children under 5 years old.Rotavirus NSP1, a 55 ku RNA binding protein, is the product of gene 5, which can subvert innate immune responses and be one of virulent determinant factors.According to the sequence in GenBank, specific primers targeting to NSP1 gene were designed and the gene was amplified by RT-PCR, following by being cloned into the pET-28a(+) vector.It showed that the full length of NSP1 gene was 1 473 bp, encoding 491 amino acids.The NSP1 shared the highest identity with WC3 strain.The recombinant protein was induced in E.coli Rosetta(DE3) by IPTG and was analyzed by SDS-PAGE and Western blotting.The results revealed that NSP1 recombinant protein existed in the form of inclusion body with the molecular weight of 55 ku.The purified recombinant protein could be recognized by His-tag antibody.This study laid the foundation for further research on the relationship between the intracytoplasmic location of NSP1 protein and its activity.

Key words: bovine rotavirus UK strain; NSP1; sequence analysis; prokaryotic expression

CLC Number:

Q786

RAN Xu-hua, CAO Si, ZHANG Yao, WEI Xiao-man, WEN Xiao-bo, NI Hong-bo, ZHU Zhan-bo. Sequence Analysis and Prokaryotic Expression of NSP1 Gene of Bovine Rotavirus UK Strain[J]. China Animal Husbandry & Veterinary Medicine, 2015, 42(5): 1104-1109.

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Sequence Analysis and Prokaryotic Expression of NSP1 Gene of Bovine Rotavirus UK Strain

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