人ZP3基因的RT-PCRcDNA克隆  

Cloning of Human ZP3 Gene cDNA by RT-PCR

作　　者：叶志华[1] 梁建庆[1] 

机构地区：[1]广东省微生物研究所

出　　处：《生物技术》2005年第4期1-4,共4页Biotechnology

基　　金：广东省科技攻关项目("重组人卵透明带-3蛋白的高效表达及其分离制备的研究";编号:2004B10401004)

摘　　要：目的:研究人ZP3基因的结构及构建人ZP3基因原核表达系统。方法:从人卵巢组织中分离出mRNA并以此作为模版,通过RT-PCR扩增出人ZP3基因cDNA片段,然后将其克隆在pUC18质粒上,并对克隆片段进行序列分析。结果:共克隆到ZP3-A(1300bp)、ZP3-B(1180bp)、ZP3-C(1200bp)和ZP3-D(1080bp)4种不同长度的人ZP3基因cDNA片段,对其中最长的ZP3-A片段的测序结果表明,它包含了人ZP3基因阅读框内的全部序列,与NCBISequenceViewer中公布的人ZP3mRNA序列(NM-007155)相比较,在1275bp长的编码区内只有一个碱基不同,两者同源性达到99.92%。结论:本研究克隆到的ZP3-AcDNA片段确是人ZP3基因无疑。Objective：To study the structure of human ZP3 gene and to consturct the prokaryotic expression system of human ZP3 gene. Method：The human ZP3 gene cDNA were amplified by RT - PCR us/rig the template mRNA isolated from the tissue of human ovary. The cDNA fragments were cloned in vector pUC18 and were then analysed by sequencing. Result： Four differential length cDNA fragments （ZP3 - A 1300 bp, ZP3 -B 1180 bp, ZP3 - C 1200 bp and ZP3 - D 1080 bp）had been cloned . The ZP3 - A fragment was the longest of them and had been sequenced.As compared with the sequences of the human ZP3 mRNA published in NCBI Sequence Viewer（NM- 007155）, the ZP3 - A cDNA fragment contained all the sequences of open reading frame of the human ZP3 gene. The homology between the two sequences were 99.92% with an only one base difference in the 1275 bp coding region. Conclusion： The ZP3 - A cDNA fragment cloned in this work was a truly fragment of the human ZP3 gene.

关 键 词：人ZP3基因 CDNA克隆 序列 

分 类 号：Q781[生物学—分子生物学] Q786