期刊论文

Construction and identification of pprM-complemented Deinococcus radiodurans strain

中文

国际放射医学核医学杂志

1673-4114

2016

40

2

115-119

10.3760/cma.j.issn.1673-4114.2016.02.006

国家自然科学基金（81272993） 湖南省研究生科研创新项目（CX2013B398）

本校为第一机构

目的 构建耐辐射奇球菌prM基因缺失回补菌株,为进一步研究PprM蛋白的功能奠定实验基础.方法 设计并合成HA-pprM基因（HA为流感病毒血凝素）全长的引物,以无突变的pGEX-6p-1-pprM载体为模板,PCR扩增出携带NdeⅠ酶切位点的HA-pprM基因全长,经NdeⅠ酶切后,与pRADK载体连接并转化大肠杆菌,经培养、提取质粒及纯化后,采用酶切鉴定以及基因测序方法确认插入序列的正确性;采用CaC12法将构建好的重组质粒pRADK-HA-pprM转化到耐辐射奇球菌pprM基因缺失的菌株中,通过Western blot验证HA-PprM融合蛋白的表达.结果 构建的重组质粒pRADK-HA-pprM经NdeⅠ酶切鉴定结果符合目的条带大小（438 bp）,测序结果显示插...

Objective To investigate the function of PprM protein by constructing a pprMcomplemented strain of Deinococcus radiodurans.Methods HA-pprM gene was amplified by PCR from the plasmid DNA of pGEX-6p-l-pprM（approximately 438 bp） and then inserted into the shuttle expression vector pRADK to construct pRADK-HA-pprM.The correctness of the inserted sequences was confirmed through agarose gel electrophoresis,restriction enzyme digestion,and gene sequencing.The recombinant plasmid was transformed into the pprM mutant strain of Deinococcus radiodurans,...