中国畜牧兽医2017，Vol.44Issue(1)：214-220,7.DOI:10.16431/j.cnki.1671-7236.2017.01.030
伽氏疏螺旋体尚志株P66基因的原核表达与鉴定
Prokaryotic Expression and Identification of P66 Gene from Borrelia garinii SZ
摘要
Abstract
In order to obtain the P66 protein from Borrelia garinii (B.garinii)SZ,the first strand cDNA was synthesized based on the total RNA extracted from B.garinii SZ,and then the targeted P66 gene was amplified by PCR.The fragment was linked into the pET-30a(+)vector,and transformed in-to Escherichia coli BL21(DE3).After identified by PCR,double restriction enzyme digestion,and nu-cleotide sequencing,the recombinant protein was expressed and purified.Polyclonal antibody was then prepared from New Zealand rabbit immunized with purified recombinant protein.The recombinant pro-tein was about 70 ku in size confirmed by SDS-PAGE,and Western blotting analysis indicated that the recombinant P66 protein could recognize the mouse monoclonal anti-His-tag,positive sera of spirochete from mouse,and anti-P66 polyclonal antibody.Additionally,the anti-P66 polyclonal antibody could rec-ognize native P66 protein.In this study,we successfully expressed the recombinant P66 protein and ob-tained the anti-P66 polyclonal antibody,which provided the foundation for further functional studies of P66 protein from B.garinii SZ.关键词
伽氏疏螺旋体尚志株/P66/原核表达/多克隆抗体Key words
Borrelia garinii SZ/P66/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
于培发,殷宏,刘志杰,牛庆丽,杨吉飞,王振国,翟斌涛,潘玉平,关贵全,罗建勋..伽氏疏螺旋体尚志株P66基因的原核表达与鉴定[J].中国畜牧兽医,2017,44(1):214-220,7.基金项目
农业科技创新工程（ASTIP）；国家肉牛牦牛产业技术体系 （）
