重组人成骨蛋白-1基因在大肠杆菌中的克隆和非诱导表达

目的　研究重组人成骨蛋白 -1(rhOP -1)在大肠杆菌中的克隆和非诱导表达。方法　利用RT -PCR技术 ,从正常胎儿肾组织总cDNA中扩增得到rhOP- 1成熟肽基因片段 ,再将rhOP- 1成熟肽基因克隆于原核融合表达载体pGEX- 4T- 2中 ,构建表达质粒pCCBC- OP- 1,并转化到大肠杆菌JM10 9(DE3)中 ,在发酵过程中不添加任何诱导剂。表达产物经SDS -PAGE分析。结果　扩增的目的基因序列与文献报道一致。表达的目的蛋白相对分子质量为 14 0 0 0 ,表达量达到菌体蛋白的 4 0 %。结论　克隆了人OP -1成熟肽基因 ,并实现了rhOP- 1的非诱导高效表达。

Gene Cloning and Non-induced Expression of Recombinant Human Osteogenic Pro-tein-1 in E. coli

Objective To clone recombinant human osteogenic protein-1(rhOP-1) gene for non-induced expression in E.coli.Methods The gene fragment encoding mature peptide of rhOP-1 was amplified from the tota l cDNA of normal fetal kidney tissue by RT-PCR and inserted into prokaryotic ex pression vector pGEX-4T-2.The constructed recombinant plasmid pCCBC/OP-1 was transformed to E.coli JM109(DE3) for non-induced expression.The expressed p roduct was identified by SDS-PAGE.Results The sequence of amplified rhOP-1 gene was identical to that reported.The expres sed product,with a relative molecular weight of 14 000,contained 40% of total so matic protein.Conclusion The gene encoding mature peptide of rhOP-1 was cloned,and rhOP-1 was highly ex pressed in E.coli without induction.