中国兽医科学2017，Vol.47Issue(10)：1275-1280,6.DOI:10.16656/j.issn.1673-4696.2017.10.012
传染性脓疱病毒112基因的原核表达及亚细胞定位
Prokaryotic expression and subcellular localization of orf virus 112 gene
摘要
Abstract
This study was designed to prokaryotically express and subcellularly localize the 112 gene of the ORFV AH-F10 strain.At first,112 gene was amplified by PCR.Then,the prokaryotic and eukaryotic expression recombinant plasmids named pET-32a-112 and pEGFP-C3-112 were successfully constructed.Subsequently,112 recombinant proteins were acquired by induction with IPTG.The expressed protein was purified and immunized with BALB/c mice to prepare for antiserum.The pEGFP-C3-112 was transfected into BHK-21 cells by Lipofectamine 2 000.Then,the subcellular localization of the expressed 112 protein was observed by inverted fluorescence microscopy.The results showed that pET-32a-112 was successfully expressed in the form of inclusion bodies in E.coli with a size of about 60 ku.The results of Western-blot demonstrated that the expressed 112 protein had good reactionogenicity with the antiserum.The results of subcellular localization showed that the 112 protein was mainly located in the cytoplasm.The present study laid a theoretical foundation for further study of thefunction of 112 protein.关键词
传染性脓疱病毒/112基因/原核表达/亚细胞定位Key words
orf virus/112 gene/prokaryotic expression/subcellular localization分类
农业科技引用本文复制引用
王勇,李永东,潘飞,杨侃侃,王元红,俞赵荣,梅跃辉,崔佣楸,苏莉,徐前明..传染性脓疱病毒112基因的原核表达及亚细胞定位[J].中国兽医科学,2017,47(10):1275-1280,6.基金项目
国家自然科学基金项目(31602063) （31602063）
安徽省自然科学基金项目(1508085QC60) （1508085QC60）
安徽农业大学稳定和引进人才科研资助项目(yj2015-16) （yj2015-16）
安徽农业大学大学生科技创新基金项目 （）
