人卵透明带重组融合基因及其表达载体的构建

目的构建人卵透明带蛋白3（zona pelludda 3，ZP3）／GST蛋白融合基因原核表达载体。方法利用PCR法扩增出去N端的信号肽和C端的跨膜区huZP3成熟蛋白编码基因。先将其克隆到pGEM-T载体，进行序列测定和分析，然后将huZP3核心DNA片断克隆入GST蛋白融合原核表达载体pGEX4T-1内。结果获得一个核苷酸长度约为1000bp的基因，与GenBank（NCBI：M60504）公布的ZP3基因序列有99％的同源性。DNA序列分析发现。有一个氨基酸出现变异。酶切鉴定后证明ZP3基因完整插入GST融合蛋白原核表达载体pGEX4T-1中。结论成功构建出含ZP3基因的GST融合蛋白原核表达载体。

Construction of recombination human zona peiiucida3/GST fusion protion gene expression vector(四)

Objective] To construct GST fusion protion gene expression vector which can express human zona peiiucida in Prokaryotic cells. Methods] The ZP3 gene was amplified by using PCR. The amplified DNA fragment was cloned into pGEM-T vector for sequencing analysis. The ZP3 gene was then subcloned into fusion protion expression vector pGEX4T-1. Result] A gene of 1 000 bp was obtained and showed a 99% homology with ZP3 gene sequence published in GenBank(NCBI: M60504). ZP3 gene was inserted into Prokaryotic expression vector pGEX4T-1 correctly. Conclusion] Having constructed ZP3 GST protion expression vector successfully.