| 摘要: |
| 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-Codon plusTM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)-Codon plusTM-RP和BL21(DE3)-Codon plusTM-RIL中获得高效表达，目的蛋白占总菌体蛋白的30％左右。 |
| 关键词: 人表皮生长因子 基因克隆 偏爱密码子 基因表达 |
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| 基金项目:国家“863”基金项目(2001AA213081)；西北农林科技大学重点专题基金项目 |
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| Gene cloning and expression of human epidermal growth factor |
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| Abstract: |
| 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 BL21(DE3)-Codon plusTM,which don't need added program to solve the coding of rare codons.pGEX-4t-1(+)-hEGF,pGEX-6p-1(+)-hEGFwas highly expressed in BL21(DE3)-Codon plusTM-RP,BL21(DE3)-Codon plusTM-RIL and the recombinant hEGF was ahout 30% of total protein measured with SDS-PAGE. |
| Key words: hEGF gene cloning rare codons gene expression |
