人角质细胞生长因子的克隆及表达

目的为获得高表达的重组人角质细胞生长因子生物工程菌。方法应用ＲＴ－ＰＣＲ技术，从人胚 胎肺成纤维细胞中，扩增出ＫＧＦ ｃＤＮＡ基因，并插入ｐＵＣ１８－Ｔ载体，构建成了重组人ＫＧＦ基因，经ＤＮＡ测序证实与 文献报道一致。将ＫＧＦ基因定向插入大肠杆菌表达载体ＰＥＴ１１ｂ内，转化大肠杆菌ＨＢ１０１。结果获得的重组子 经ＩＰＴＧ诱导，在相对分子质量为１９０００处表达重组蛋白，约占菌体蛋白的１０％。结论建立了制备重组人ＫＧＦ表 达系统，为今后进一步研究ＫＧＦ的生物学功能奠定可靠的物质基础。

Cloning and Expression of Recombinant Human Keratinocyte Growth Factor(KGF)

Objective To obtain the gene engineering bacterial strain expressing human keratinocyte growth factor.Mehtods The cDNA gene encoding keratinocyte growth factor(KGF)was amplified from human embryonic fibroblast by RT-PCR and inserted into pUC18-T vector.The recombinant KGF gene was directly inserted into expression plasmid pET11b and transformed to E.coli HB 101.Results DNA sequencing result of the recombinant KGF gene was consistent with that reported. About 10% of somatic proteins were expressed, and the relative molecular weight of the expressed product was 19000. Conclusion A expression system of recombinant human KGF was established. It Provided a reliable matter basis for the further study on the biological characteristics of KGF.