CAJ | 학술논문

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
목적　탐토인간세포인자적결구여공능급기재진핵세포중적표체여조공적궤제，수선극륭료인간세포인자전장cDNA。방법　용RPMI1640，체적분수위10%적소우혈청배양HepG2세포，추제RNA，행RT-PCR，순화PCR산물，여pGEM-T극륭재체련접，전화，포판，사선양성극륭，매절감정병진행서렬측정。결과　통과매절감정병진행서렬측정，성공지획득료인간세포인자전장cDNA적기인극륭。결론 　인간세포인자전장cDNA적기인극륭적구건성공，위기결구여공능적연구급기재진핵세포중적표체여조공적연구제공료일정적물질기출
Objective To study the expression and regulation of human stem cell factor in eukaryotic cell, its full-length cDNA 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.