人表皮生长因子的基因克隆与原核表达

Gene cloning and expression of human epidermal growth factor

采用RT-PCR方法从人胎盘组织中扩增出hEGF基因,将其分别连入表达载体pGEX-4t-1(+)和pGEX-6p-1(+)质粒中,并转化入BL21-CodonplusTM中,解决了大肠杆菌密码子偏爱性问题,不需附加程序就可以编码稀有密码子的重组基因。SDS-PAGE结果证明,pGEX-4t-1(+)-hEGF和pGEX-6p-1(+)-hEGF分别在大肠杆菌BL21(DE3)-CodonplusTM-RP和BL21(DE3)-CodonplusTM-RIL中获得高效表达,目的蛋白占总菌体蛋白的30%左右。

Human EGF gene was successfully amplified from human placenta with RT-PCR method,then cloned it into expression vector:pGEX-4t-1(+)and pGEX-6p-1(+) and shift them to BL_(21)(DE3) -Codon plus^(TM),which don't need added program to solve the coding of rare codons.pGEX-4t-1(+)-hEGF,pGEX-6p-1(+)-hEGFwas highly expressed in BL_(21)(DE3) -Codon plus^(TM) -RP,BL_(21)(DE3) -Codon plus^(TM) -RIL and the recombinant hEGF was ahout 30% of total protein measured with SDS-PAGE.