GDF5基因启动子表达载体的构建

Construction and expression of vector driven by GDF5 promoter

目的构建含人生长分化因子5(GDF5)基因启动子表达载体pGL3-GDF5,为研究GDF5在骨骼、肌肉发育的作用机制提供实验基础。方法扩增人GDF5基因启动子序列,克隆至pGL3-Basic表达载体并转化入大肠杆菌DH10b,双酶切鉴定并测序。结果 PCR扩增后的产物在约854 bp处出现明显的特异性条带,酶切鉴定结果与理论预测值一致,测序未出现碱基突变。结论成功构建了含GDF5基因启动子的表达载体。

Objective To construct the expression vector containing growth differentiation factor 5（GDF5） promoter pGL3-GDF5,providing experimental basis for the studying of GDF5 in the mechanism of development of bone and muscle.Methods GDF5 was amplified with the human genome DNA by PCR,the segment was cloned into the pGL3-Basic vector and transformed into E.Coli,which was detected by endonuclease and sequenced.Results The PCR amplified products appeared specific band at 854 bp site.The sequenced segment（854 bp） in the recombinants was identical to that GenBank had reported and the segment was inserted in right direction.Conclusion The expression vector pGL3-GDF5 is successfully constructed.