小鼠胰岛素样生长因子结合蛋白7原核表达载体的构建

Cloning and Expression of Mouse IGFBP7 cDNA in E. coli

胰岛素样生长因子结合蛋白 7(简称IGFBP7)广泛存在于多种正常的组织中 ,但在相应的肿瘤细胞中的含量极微。试验表明 ,它在肿瘤发生的不同阶段抑制其生长。采用RT PCR和Nest PCR方法 ,从正常的小鼠脾脏中扩增得到IGFBP7cDNA片段 ,测序正确后 。

The full length cDNA encoding mouse insulin like growth factor binding protein 7 (mIGFBP7) was amplified by reverse transcription polymerase chain reaction (RT PCR)and nest polymerase chain reaction (Nest PCR) from the spleen of a normal mouse (BALB/C).After purifying and sequencing ,the product was inserted into the prokaryotic fusion expression vector PRSETC ,an IGFBP7 gene recombinant expression vector plasmid was constructed ,then the plasmid was transfected into E.coli BL21DE3.The SDS PAGE showed that E.coli BL21DE3 expressed a His6 fusion protein containing pRIGFBP7.