人层粘连蛋白α4链LG3组件的克隆、测序及在原核细胞中的表达

Cloning,Sequencing and Prokaryotic Expression of Human Laminin alpha 4 Chain LG3 Module

用Trizol法提取胎盘组织总RNA ,通过RT PCR方法获得人层粘连蛋白α4链LG3组件的特异扩增产物。将PCR产物克隆入PMD 1 8T载体中 ,PCR与酶切鉴定正确后进行序列测定。用测序正确的克隆片段与原核表达载体PET 2 8a进行体外重组 ,构建了LG3的原核表达载体 ,并在BL2 1 (DE3)菌株中得到了表达。

Total RNA was extracted from placenta by Trizol reagent,and an unique RT-PCR fragment of human laminin alpha 4 chain LG3 module was obtained.The PCR product then was cloned into PMD-18T vector and sequenced after it was identified by enzyme digestion and PCR amplification.Recombinant LG3 prokaryotic expression vector was constructed through recombination of PET-28a vector and certificated by sequencing,and the finally recombinant LG3 module was got expressed in E.coli BL21(DE3) strain. ;