【作者】 程昀静； 张利仙； 陶俊； 段纲； 艾军； 龙云凤； 董俊； 严树发； 周晓黎；

【Author】 CHENG Yun-Jing ZHANG Li-Xian TAO Jun DUAN Gang,AI Jun~* LONG Yun-Feng DONG Jun YAN Shu-Fa ZHOU Xiao-Li~* (1.College of Animal Science and Technology,Yunnan Agricultural University,Kunming,650201;2.kunming University of Scienceand Technology,Kunming,650224;3.Kunming Medical University,Kunming,650031;4.Yunnan Entry-Exit Inspection and Quarantine Burea,Kunming,650228)

【机构】 云南农业大学动物科学技术学院； 昆明理工大学； 昆明医学院； 云南出入境检验检疫局；

【摘要】 利用PCR扩增SRV-2型病毒株的P27基因,将扩增片段克隆入原核表达载体pBAD/Thio-TOPO,通过序列分析证实该片段与猴D型逆转录病毒SRV-2株P27基因序列一致。将阳性重组质粒转化至大肠杆菌TOPO10中,用阿拉伯糖诱导表达,表达的融合蛋白进行SDS-PAGE和WesternBlot分析,并用proBondTM柱进行纯化。结果表明,表达的融合蛋白分子量约为50KDa,其大小与理论值相符。更多还原

【Abstract】 The simian type-D retrovirus p27 gene was amplified by PCR from SRV-2 strain.The PCR fragment was cloned into prokaryotic expression vector Pbad/Thio-TOPO vector and sequenced.The amplified p27 sequence was identical to that of SRV-2 strain.The recombinant press vector pressed in the host cell TOPO10 under Arabinose induction.The expressed recombinant product was identified by SDS- PAGE and Western Blot,and purified by proBondTM column under native conditions.The results demonstrated that recombinant protein had a molecular weight of 50 KDa and its size in line with expectations.更多还原