【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。更多还原

【Abstract】 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 using 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.更多还原