骨形成蛋白-2和转化生长因子-β1双基因真核表达载体的构建

Construction of bicistronic eukaryotic expression plasmid containing BMP-2 and TGF-β1 genes

目的构建骨形成蛋白（BMP）-2、转化生长因子（TGF）-β1双基因真核表达载体。方法以pGEM-T-BMP-2及pGEM-T-TGF-β1两个质粒为模板，分别用引入新的酶切位点引物，PCR扩增出长度为1188bp的BMP-2和1173bp的TGF-β1两个目基因片段；将其分别定向连入真核双基因表达载体pIRES；酶切分析及序列测定重组子。结果双酶切分析电泳可见长度为1188bp的BMP-2条带和1173bp的TGF-β1条带，核酸序列测定证实重组质粒pIRES-BMP-2-TGF-β1构建正确。结论成功构建了BMP-2及TGF-β1双基因真核表达载体。

Objective To construct a bicistronic eukaryotic expression plasmid consisting of BMP-2 and TGF-β1 target genes. Methods Firstly,the DNA fragments of 1188 bp BMP-2 and 1173 bp TGF-β1 gene were obtained from pGEM-T-BMP-2 and pGEM-T-TGF-β1 plasmids by PCR respectively. Then they were inserted into bicistronic eukaryotic expression plasmid vector plRES. The inserted target genes in the plasmid were verified by restriction enzyme digestion and nucleotide sequencing. Results The direction and sequences of the new bicistronic eukaryotic expression plasmid pIRES-BMP-2-TGF-β1 with 1188 bp BMP-2 and 1173 bp TGF-β1 were correct. Conclusion The bicistronic eukaryotic expression plasmid is constructed successfully.