人TAT-PDX-1融合蛋白原核表达载体的构建  

Construction of prokaryotic expression vector of human TAT-PDX-1 fusion protein

作　　者：苗成[1] 张建璞[1] 

机构地区：[1]新乡市中心医院神经内科,河南新乡453000

出　　处：《新乡医学院学报》2011年第3期304-306,共3页Journal of Xinxiang Medical University

摘　　要：目的构建人TAT-PDX-1融合蛋白原核表达载体。方法以人胰腺癌(PANC)-1细胞株cDNA为模板,采用反转录聚合酶链式反应(RT-PCR)扩增胰腺十二指肠同源异型盒基因-1(PDX-1)蛋白编码的全部序列,克隆入原核表达载体pET28a中,经限制性内切酶HindIII和BamHI双酶切及DNA序列分析重组质粒的目的基因。结果 RT-PCR扩增产物的特异性片段长度为885 bp,以此构建的重组质粒pET28a-TAT-PDX-1经HindIII和BamHI双酶切后显示5 900 bp和885 bp左右的2条片段,测序结果与Genbank中的人PDX-1基因cDNA序列一致。结论成功构建了人TAT-PDX-1融合蛋白原核表达载体。Objective To construct a prokaryotic expression vector of human TAT-PDX-1 fusion protein.Methods cDNA of human pancreatic（PANC）-1 cell line as a template,all coding sequence of PDX-1 protein were amplified by reverse transcription polymerase chain reaction（RT-PCR）.The fragment was inserted into prokaryotic expression vector pET28a plasmid.Target genes of recombinant plasmid were analyzed by the restriction enzyme HindIII and BamHI double digestion and DNA sequencing.Results The length of specific fragment applied by RT-PCR was 885 bp,and after the pET28a-TAT-PDX-1 of recombinant plasmid digested by HindIII and BamHI double digestion displayed two fragments of 5 900 bp and 885 bp.The sequencing results consistent with cDNA sequence of human PDX-1 gene in Genbank.Conclusion The prokaryotic expression vector of human TAT-PDX-1 fusion protein is successfully constructed.

关 键 词：原核表达 蛋白转导域 PDX-1 反式激活蛋白 

分 类 号：Q756[生物学—分子生物学]