期刊论文

中文

中华实验和临床病毒学杂志

1003-9279

2015

29

1

83-86

10.3760/cma.j.issn.1003-9279.2015.01.028

国家科技重大专项（2013ZX10004001）；

本校为第一机构

目的 构建表达IL-2的重组非复制型痘苗病毒株并体外检测其生物活性.方法 以非复制型痘苗病毒天坛株为载体,通过同源重组、蓝白斑纯化筛选表达IL-2的非复制型重组痘苗病毒rNTV1175IL2H6LacZ,并通过PCR对其进行IL-2基因插入鉴定,Western Blot鉴定IL-2的表达,同时对其进行体外生物活性检测.结果 经PCR鉴定,痘苗病毒TK区7.5启动子下正确插入了IL-2基因511bp,Western Blot结果表明其表达IL-2蛋白相对分子质量大小为15000,与标准品IL-2一致.rNTV1175 IL2H6LacZ分泌表达在细胞培养液中的IL-2能使IL-2依赖CTLL-2细胞株生长,MTT法测得重组病毒表达在细胞培养液中的IL-2生物活性值为8.79 IU/ml.结论 成功构...

Objective To generate a recombinant non-replicating vaccinia virus expressing IL-2 and detect its bioactivity in vitro.Methods Based on the non-replicating vaccinia virus （Tiantan strain） vectors,recombinant virus rNTV1175IL2H6LacZ which expressed IL-2 was constructed through homologous recombination and purified by blue-white screening.Then objective gene and protein expression was identified and the bioactivity of objective protein in vitro was detected.Results Recombinant virus was identified by PCR and Western Blot indicating that IL-2 ge...