安徽农业大学学报2017，Vol.44Issue(5)：780-783,4.DOI:10.13610/j.cnki.1672-352x.20171017.025
口疮病毒O2O基因的克隆、表达及多抗制备
Cloning and expression of the Orf virus O2O gene and its polyclonal antibody preparation
摘要
Abstract
Orf is an infectious disease caused by off virus (ORFV).Animals can be infected by ORFV through contact and it is preferred to epithelial infection.The ORFV O2O gene is an important enzyme regulating protein activity of the virus.In this study,specific PCR primers were designed according to the sequence of the Off O2O gene in GenBank.After the viral DNA was extracted from ORFV infected clinical samples,The Off O2O gene was amplified by PCR and subcloned to pET-32a prokaryotic expression vector.The recombinant plasmid was identified and named as pET-32a-ORF-O2O,which was then transformed into E.coli competent cell BL21 and induced by IPTG.The expressed proteins were identified by SDS-PAGE and Western blot.The ORF O2O protein was then purified and immunized to BALB/c mice for antiserum preparation.The results of SDS-PAGE showed that the Orf O2O gene was correctly expressed in vitro.The protein size was about 37 kDa.The western-blot analysis showed that the prepared antiserum reacted specifically with the ORF O2O protein.关键词
羊口疮病毒/O2O基因/克隆/原核表达/多克隆抗体Key words
ORFV/O2O gene/clone/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
王勇,徐前明,蒋书东,赵玉洁,杨侃侃,殷冬冬,白彩霞,潘飞,梅跃辉,颜秀梅,王君..口疮病毒O2O基因的克隆、表达及多抗制备[J].安徽农业大学学报,2017,44(5):780-783,4.基金项目
国家自然科学基金(31602063),安徽省自然科学基金(1508085QC60),安徽农业大学稳定和引进人才科研项目(yj2015-16)和安徽农业大学大学生科技创新基金项目共同资助. （31602063）
