汉滩病毒糖蛋白G1 G2和核蛋白NP在痘苗病毒天坛株中的联合表达  

CO-EXPRESSION OF HANTAAN VIRUS GLYCOPROTEINS AND NP IN VACCINIA VIRUS TIANTAN STRAIN

作　　者：姚树元[1] 杭长寿[1] 邱建明[1] 马章亮[1] 李德新[1] 薛颖[1] 李川[1] 宋干[1] 

机构地区：[1]中国预防医学科学院病毒学研究所

出　　处：《病毒学报》1998年第2期121-125,共5页Chinese Journal of Virology

摘　　要：将汉滩病毒７６／１１８株Ｍ、Ｓ片段分别插入转染质粒ｐＪＳＢ１１７５的Ｐ１１和Ｐ７．５启动子下游，构建成重组质粒ｐＪＳＢ１１Ｍ７．５Ｓ。采用Ｌｉｐｏｆｅｃｔｉｎ转染技术将重组质粒分别与痘苗病毒天坛株ＴＫ＋ｖｖ和表达Ｓ片段的重组痘苗病毒ｖＪＳＡ１１７５Ｓ进行共转染，免疫酶斑和蓝白斑法筛选并纯化，得到了重组病毒ｖＪＳＢ１１Ｍ７５Ｓ。Ｂｕｄｒ的选择压力试验表明，外源基因确实插入在ＴＫ基因区；ＰＣＲ及打点杂交证实了两个片段的插入；重组痘苗病毒基因组的部分序列分析显示，两个片段与各自的启动子连接正确；间接免疫荧光及放免沉淀证明了糖蛋白Ｇ１、Ｇ２和核蛋白ＮＰ的表达。By replacing LacZ gene with HTNV M genome segment cDNA from a recombined vaccinia virus that expressed HTNV NP and LacZ,we obtained a vaccinia virus recombinant.M and S genome segments of HTNV were separately controlled by promoter P11 and P7.5 in this recombinant,glycoproteins and NP were co-expressed in the same virus.The existence of both two segments was determined by PCR and Dot blotting,the expression of glycoproteins and NP also could be proved by IFA and RIP. Neutralizing antibody could be testified with rabbit’s anti-sera induced by the recombinant.It showed glycoproteins were actually expressed by this recombinant vaccinia virus.

关 键 词：汉坦病毒 重组痘苗病毒 基因表达 糖蛋白 核蛋白 

分 类 号：R373.32[医药卫生—病原生物学] R784[医药卫生—基础医学]