重组人骨形成蛋白-3的克隆及其在大肠杆菌中的表达

Cloning and Expression of rhBMP 3 in Escherichia coli

Here is shown the cloning and expression of BMP 3 cDNA in E.coli .Utilizing specific primers designed according to published sequences,a 0 3kb band was assessed in various human tissues,namely brain,liver,thymus,spleen,placenta and testis,as well as human neuroblastoma SK cells by RT PCR.The 0 3kb fragment was inserted into pCRII vector and the sequencing result identified it the expected 288 bp BMP 3 cDNA.The expression of this BMP gene fragment was acquaired by ligating it with pGEX 2T and transforming E.coli .The transformed E.coli yielded high level of BMP 3 GST fusion proteins.Furthermore,the activities of the fusion proteins was domestrated by Western blotting.