人干细胞因子全长cDNA的基因扩增与克隆

目的 探讨人干细胞因子的结构与功能及其在真核细胞中的表达与调控的机制，首先克隆了人干细胞因子全长cDNA。方法 用RPMI1640，体积分数为10％的小牛血清培养HepG2细胞，抽提RNA，行RT－PCR，纯化PCR产物，与pGEM-T克隆载体连接，转化、铺板，筛选阳性克隆，酶切鉴定并进行序列测定。结果 通过酶切鉴定并进行序列测定，成功地获得了人干细胞因子全长cDNA的基因克隆。结论 人干细胞因子全长cDNA的基因克隆的构建成功，为其结构与功能的研究及其在真核细胞中的表达与调控的研究提供了一定的物质基础。

Amplification and cloning of human stem cell factor full-length cDNA by RT-PCR

Objective To study the expression and regulation of human stem cell factor in eukaryotic cell, its full - length cD-NA was amplified by RT - PCR and its cloning vector was constructed. Methods HepG2 cells were cultured in RPMI1640 containing 10% bovine serum, and the cultured cells were harvested and their RNA was extracted;The 1.14 kb cDNA was amplified by RT - PCR, and the full - length cDNA fragment was inserted in the pGEM - T vector. Results The recombinant plasmid was cleaved with restrictive endonuclease and sequencing result showed that the cloning vector was successfully created. Conclusion The recombinant construct of the full - length cDNA provids available conditions for further study on expression and regulation of stem cell factor.