人胰岛素样生长因子-1基因克隆及其真核表达载体的构建

Clone of human IGF-1 gene and its construction with eukaryotic expression vector

目的:从胎儿肝组织中克隆人胰岛素样生长因子-1(IGF-1)基因,并构建真核表达载体pCI-neo/hIGF-1。方法:提取胎儿肝总RNA,RT-PCR法扩增IGF-1 cDNA片段,重组于pUCM-T载体,测序正确后构建表达载体。结果:扩增得到710bp带有Kozak序列的IGF-1 cDNA片段,成功构建了真核表达载体pCI-neo/hIGF-1。结论:通过RT-PCR的方法克隆了人IGF-1 cDNA,并成功构建了真核表达载体pCI-neo/hIGF-1,为IGF-1基因治疗糖尿病创造了条件。

Objective： To clone human IGF-1 gene from the fetus liver and construct eukaryotic expression vector pCI-neo/ hIGF-I. Methods： The total RNA of the fetus liver were extracted; IGF-1 cDNA fragment was amplified with RT-PCR and inserted into pUCM-T vector, and then the expression vector was constructed. Results： 710 bp IGF-1 cDNA fragment with Kozak sequence was obtained by amplification and the pCI-neo/hIGF-1 eukaryotic expression vector was successfully con- structed. Conclusion： Human IGF-1 cDNA was cloned by RT-PCR and eukaryotic expression vector containing hlGF-1 was successfully constructed, providing a possibility for curing diabetes by IGF-I gene.