中国畜牧兽医2016，Vol.43Issue(9)：2291-2295,5.DOI:10.16431/j.cnki.1671-7236.2016.09.011
羊口疮病毒 ORF 059基因的真核表达研究
Study on Eukaryotic Expression of ORF059 Gene of Orf Virus
摘要
Abstract
The study was aimed to clonie and express ORF 059 gene of Orf virus (ORFV).The ge-nome of ORFV HCE attenuated vaccine strain was extracted and used as the template.Based on the referenced gene sequences of ORF 059 gene on GenBank,the specific primers were designed, the fragment of ORF 059 gene was amplified by PCR.After being purified,ORF 059 gene was in-serted into pEGFP-N1 vector to construct recombinant plasmid pEGFP-ORF059-N1.After being sequenced,the recombinant plasmid pEGFP-ORF059-N1 was transfected into NIH3T3 cells with liposomes.The expression of fusion protein was identified by the fluorescence microscope obser-vation and Western blotting.The results showed that the ORF 059 gene was 1 005 bp.The recom-binant plasmid pEGFP-ORF059-N1 was constructed correctly.Green fluorescence could be ob-served in NIH3T3 cells transfected by pEGFP-ORF059-N1 under the fluorescence microscope. The expressed fusion protein was about 64 ku.The results laid a foundation for the further re-search on the mechanism of ORF059.关键词
羊口疮病毒/ORF059 基因/真核表达/NIH3T3 细胞Key words
ORFV/ORF 059 gene/eukaryotic expression/NIH3T3 cell分类
生物科学引用本文复制引用
朱姝,杜丽,庞峰,李国华,李亚颖,彭冬梅,杨小健,聂鑫,曹瑞勇,王凤阳..羊口疮病毒 ORF 059基因的真核表达研究[J].中国畜牧兽医,2016,43(9):2291-2295,5.基金项目
海南省重大科技计划项目(ZDKJ2016017-01) （ZDKJ2016017-01）
