GAPB基因原核表达载体的构建  

Construction of Glyceraldehyde-3-phosphate dehydrogenase B subunit gene vector expressing in prokaryotic system

作　　者：田霞[1] 代其林[1] 龚元亚[1] 孙英坤[1] 谢琳[1] 杨娟[1] 王劲[1,2] 

机构地区：[1]西南科技大学植物生物技术研究中心,四川绵阳621010 [2]中国农业科学院生物技术研究所,北京100081

出　　处：《南方农业》2011年第3期1-4,共4页South China Agriculture

基　　金：国家转基因生物新品种培育重大专项(2009ZX08009-091B);国家自然科学基金(30871555);教育部新世纪优秀人才支持计划(NCET-08-0940);四川省教育厅科技项目(09ZA034)

摘　　要：以拟南芥cDNA为模板,用PCR扩增出GAPB的基因全长,然后将GAPB基因片段连接到PET28a(+)载体上,构建重组质粒并转化大肠杆菌DH5α,经菌落PCR和酶切鉴定筛选出阳性克隆,测序正确后,再将阳性克隆的质粒转化大肠杆菌BL21(DE3)。结果表明:成功构建了原核表达载体PET28a(+)-GAPB,为后续的GAPB融合蛋白的表达以及纯化奠定了基础。In this study,GAPB DNA fragment was amplified by polymerase chain reaction from Arabidopsis cDNA template.The GAPB DNA fragment was then connected into plasmid PET28a（＋）,and was transformed into E.coli DH5α.The positive clones were screened by colony PCR and enzyme digested,which were transformed into E.coli BL21 DE3 star plyss after sequencing.It was concluded that the prokaryotic expression plasmid PET28a（＋）GAPB was built successfully,which provided a material foundation for expression and purification of GAPB fusion protein.

关 键 词：GAPB 大肠杆菌BL21 PET28a(+) 融合蛋白 

分 类 号：Q946[生物学—植物学]