人促血小板生成素(TPO)原核表达载体pQE30-TPO的构建

目的：获得人血小板生成素全长cDNA克隆，并构建重组表达载体pQE30-TPO。方法：利用RTPCR技术从人胎肝细胞mRNA中扩增目的基因，将扩增产物克隆至pMD18-T载体，经菌落PCR鉴定后，DNA序列分析重组质粒pMD18－T－TPO。构建并鉴定原核表达载体pQE30-TPO。结果：构建了重组克隆载体pMD18-T-TPO和重组表达载体pQE30-TPO，序列分析表明，获得的TPO cDNA序列与国内外报道的人促血小板生成素核苷酸序列完全一致。结论：成功构建了重组表达载体pQE30-TPO，为TPO的进一步研究奠定了基础。

Construction of human thrombopoietin(TPO) prokaryotic expression vector pQE30-TPO

Objective:To clone human thrombopoietin cDNA and to construct human TPO prokary-otic expression vector.Method:RT-PCR was used to amplify the target gene with total RNA from human fetal liver cell.The gene was inserted into pMD18-T,identified by bacteria colonies PCR,then sequenced.After that,the prokaryotic expression vector pQE30-TPO was constructed and iden tified.Results:Clonal re combi nant of pMD18-T-TPO and prokaryotic expression vector pQE30-TPO were constructed.The ob-tained TPO cDNA sequence is the same as what had been reported previously.Conclusion:Prokaryotic expression vector pQE30-TPO was successfully constructed.