人重组内皮抑素基因的克隆与融合表达

CIONING AND FUSION EXPRESSION OF RECOMBINANT HUMAN ENDOSTATIN GENE

目的构建人内皮抑素(endostatin)融合蛋白表达载体,并在大肠杆菌中表达。方法用反转录多聚酶链反应(RT-PCR)得到人内皮抑素全基因,连接PGEM-T载体,测序确认,定向克隆重组入pTRX表达载体,转化大肠杆菌BL21(DE3),IPTG诱导表达。结果经SDS-PAGE分析,出现特异性蛋白质条带,大小与预期相符合。结论人内皮抑素基因在原核细胞表达载体pTRX中获得成功表达,为应用内皮抑素进行肿瘤的抗血管生成治疗奠定了基础。

Objective To clone human endostatin gene into fusion expression vector and to express in E.coli BL21(DE3). Methods Human endostatin gene was acquired by means of RT-PCR, and was cloned into PGEM-T vector and its sequence was identified. The gene was inserted into fusion expression vector pTRX, and transformed in E.coli BL21(DE3),then endostatin was expressed in E.coli by IPTG inducement. Results SDS-PAGE analysis showed a specific protein strip which was identical to the expected one. Conclusion Human endostatin gene was successfully expressed in E.coli. The experiment lays foundation for antiangiogenesis therapy with endostatin gene.