BMP2转基因载体的构建及在大肠杆菌DH-5α中的诱导表达

Construction of pcDNA3-BMP2 and Induced Expression in E. coli DH-5α

BMP2基因来源于pSP6-BMP2质粒,用pcDNA3作载体,构建BMP2转基因载体并在大肠杆菌DH-5α中的诱导表达。用HindⅢ与XbaI双酶切pSP6-BMP2与pcDNA3质粒,回收BMP2基因片段与pcDNA3载体,用连接酶连接后转化JM109,提质粒后酶切鉴定;把构建好的载体转化DH-5α细菌,并诱导表达,用SDS-PAGE电泳鉴定有无表达,用Western Blot鉴定表达蛋白。pcDNA3-BMP2载体酶切鉴定与预期片段相符,SDS-PAGE显示有BMP2蛋白表达;Western Blot证明表达蛋白有免疫原活性。成功地构建了表明pcDNA3-BMP2转基因载体并在大肠杆菌DH-5α中诱导表达了BMP蛋白。

BMP2 gene fragment was from pSP6-BMP2, pcDNA3-BMP2 vector was constructed with pcDNA3 ,and expressed in E. coil DH-5α. pSP6-BMP2 and pcDNA3 were cut by HindⅢ and XbaI, extracted BMP2 fragment and pcD-NA3 vector, liangased BMP2 and pcDNA3 together, and then transfected into JM109,transferred plasmid was extracted out and identified by enzyme cut . Constructed vector was transferred into E. coli DH-5α. BMP2 protein was induced to express in E .coli DH-5α and identified by SDS-PAGE and Western blot. Enzyme cut confirmed pcDNA3-BMP2 vector was constructed successful. Result of SDS-PAGE identified with BMP2 expression in E. coli DH-5α transferred by pcD-NA3-BMP2 . Result of Western blot identified expressed protein in E.coli. DH-5α was with acitivity.