BMP2和TGFβ3双基因真核表达载体的构建

Construction of bicistronic eukaryotic expression plasmid containing BMP2 and TGFβ3 genes

[目的]构建骨形态发生蛋白BMP2和转化生长因子TGFβ3双基因真核表达载体。[方法]从人胚胎组织提取总RNA,反转录成cDNA,以pGEMT/BMP2和反转录的cDNA为模板,PCR扩增出BMP2和TGFβ3两个基因全长,将两个基因片段分别定向连入双基因真核表达载体pIRES;用酶切的方法筛选出阳性重组质粒,并进行测序鉴定。[结果]酶切鉴定证明已将BMP2和TGFβ3两个基因连入载体中,测序结果完全正确。[结论]成功构建pIRES-BMP2/TGFβ3双基因真核表达载体。

[Objective]To construct a bicistronic eukayotic expression plasmid consisting of BMP2 and TGFβ3 target genes.[Method]The DNA fragment of BMP2 and TGFβ3 genes were obtained from pGEMT/BMP2 plasmid and human embryonal tissue by PCR,respectively.They were inserted into bicistronic eukaryotic expression plasmid vector pIRES.The inserted target genes in the plasmid were verified by restriction enzyme digestion and nucleotide sequencing.[Result]The direction and sequences of the new bicistronic eukaryotic expression plasmid pIRES-BMP2-TGFβ3 were correct.[Conclusion]The bicistronic eukaryotic expression plasmid has been constructed successfully.