猪瘟病毒E2基因pGEME2重组质粒仍具有α互补作用

猪瘟病毒E2基因pGEME2重组质粒仍具有α互补作用 pGEM Plasmid Containing E2 Gene of CSFV Still Capable of β-galactosidase α-complementation 王宁 WANG Ning 1 first-author 傅烈振 FU Lie\|zhen 1 张楚瑜 ZHANG Chu\|yu 1 朱燕 Zhu Yan 1 武汉大学生命科学院,武汉,430072 武汉大学生命科学院,武汉,430072 College of Life Sciences, Wuhan University College of Life Sciences, Wuhan University 在利用pGEM-T载体克隆猪瘟病毒石门株E2 基因中发现,E2 基因以与lacZ相同方向插入时,重组质粒仍具有α互补作用,呈典型的蓝色菌落,而以与lacZ基因相反方向插入时,重组质粒无α互补作用,呈白色菌落. 经E2 蛋白检测、序列分析、重组质粒缺失和插入等研究,结果表明pGEME2 的α互补作用产生机制不同于目前人们已知的机制. 它由外源E2 基因内部起始合成新的α肽,从而赋于其α互补作用. 此结果表明利用α互补作用并不能筛选到全部克隆. The amplified E2 gene fragment was inserted into pGEM T vector. When E2 gene was inserted in the same orientation as lacZ, the recombinant plasmid pGEME2 was still capable of β galactosidase α complementation. When E2 gene was inserted in reverse orientation, the recombinant plasmid had no capability of α complementation, experiments indicated that α complementation of pGEME2 was achieved by a synthesis of a new α donor starting from interior E2 gene.\; 猪瘟病毒石门株 pGEM-T E2基因 α互补作用 SD序列 Key words: classical swine fever virus pGEM T vector α complementation SD Sequence Q785;S852.65 国家攀登计划B类资助!（85-41） 1999-06-01 2021-04-01 6