猪水泡病病毒VP1基因的克隆和表达.pdf

第21 卷4 期 中 国 病 毒 学 21 （4 ）：385-389 2006 年7 月 VIROLOGICA SINICA July 2006 猪水泡病病毒VP1 基因的克隆和表达* 1 2** 2 1 2 钟金栋 ，花群义 ，肖荣海 ，夏雪山 ，杨云庆 ， 2 2 3 周晓黎 ，董 俊 ，邓祖洪 （1.昆明理工大学生化学院,云南昆明650224 ；2.云南出入境检验检疫局技术中心，云南昆明650228 ；3.西双版纳兽医站,云南景洪666100 ） Cloning and Expression of VP1 Gene of Swine Vesicular Disease Virus 1 2** 2 1 ZHONG Jin-dong ，HUA Qun-yi ，XIAO Rong-hai ，XIA Xue-shan ， 2 2 2 3 YANG Yun-qing ，ZHOU Xiao-li ，DONG-jun ，DEN Zu-hong (1. Faculty of Bioengineering and Chemical Engineering, Kunming University of Science and Technology, Kunming 650224, China; 2. Technology Center of Yunnan Entry-Exit Inspection and Quarantine Bureau, Kunming 650228, China; 3. Xishuangbanna Veterinary Station, Jinghong 666100, China ) Abstract ：The VP1 gene of Swine vesicular disease virus (SVDV) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) yielding a product of 849bp cDNA fragment. Using T-A cloning technique, the PCR product was cloned into pMD18-T vector. The purified VP1 gene was subcloned into the pBAD/Thio TOPO vector and the plasmid was identified by PCR. It was sequenced to confirm the authenticity of the sequence and orientations. SDSand Western blotting revealed that the VP1 protein was expressed in Escherichia coli LGM194 at a high level and the recombinant fusion protein containe