Role of mouse hepatitis coronavirus envelope protein transmembrane domain.
Role of mouse hepatitis coronavirus envelope protein transmembrane domain.
复制标题
小鼠肝炎冠状病毒包膜蛋白跨膜结构域的作用。
DOI：
10.1007/978-0-387-33012-9_32
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发表时间：
2006
影响因子：
--
通讯作者：
Hogue,BrendaG
中科院分区：
文献类型：
--
作者：
Ye,Ye;Hogue,BrendaG
2.材料和方法使用全质粒PCR方案，用含有所需突变的一对引物产生丙氨酸插入突变体。将含有E基因的质粒用作PCR的模板。在确认引入的突变后，将该基因亚克隆到MHV-CoV A59感染性克隆的G亚克隆中，该感染性克隆是七个cDNA片段（A-G）系统的一部分，由查佩尔山的北卡罗来纳州的Ralph Baric博士友情提供。12使用基本上如所述的方案组装所有全长cDNA克隆并转录RNA。电穿孔后，通过噬斑纯化回收突变病毒。多发斑块
2. MATERIALS AND METHODSAlanine insertion mutants were created using a whole plasmid PCR protocol with a pair of primers containing the desired mutations. A plasmid containing the E gene was used as the template for PCR. After confirmation of the introduced mutations, the gene was subcloned into the G subclone of the MHV-CoV A59 infectious clone that is part of the seven cDNA fragment (A–G) system, kindly provided by Dr. Ralph Baric at the University of North Carolina at Chapel Hill. 12 All full-length cDNA clones were assembled and RNAs were transcribed using a protocol basically as described. 12, 13 After electroporation, mutant viruses were recovered by plaque purification. Multiple plaques