RP2突变体RP2-dser6的克隆和表达

Cloning and Expression of Mutant RP2-dser6 Gene

视网膜色素变性 (RetinitsPigmentosa ,RP)是遗传性视觉损害以致失明的最常见原因之一 ,RP2是最近定位克隆的一个视网膜色素变性基因。为研究突变RP2蛋白与正常RP2蛋白的空间结构的差异及其对功能的影响 ,用突变的寡聚核苷酸引物自重组质粒 pETRP2PCR扩增得到突变的RP2 dser6基因 ,克隆至 pET11C载体中 ,转化大肠杆菌BL2 1菌株。重组子经序列分析鉴定后 ,在 37℃诱导RP2 dser6突变蛋白表达 ,5mmol/LIPTG诱导 8小时表达量约为 7 8%。所得结果为进一步研究RP致病机理奠定了基础。

WT5BZ]RP2 is an X linked retinitis pigmentosa gene newly discovered by positional cloning, A mutant of RP2 gene, named RP2 dSer6, was obtained by PCR from recombinant plasmid pETRP2 and cloned into pET11c to construct pETRP2 dSer6, which was sequenced and used to transform BL21 strain. The expression ratio of mutant RP2 dSer6 protein was approximately 7.8 % at 37℃ under the induction of 5 mmol / L of IPTG after 8 h.