人PLAGL2基因表达载体质粒的构建

The construction of human PLAGL2 gene recombinant DNA plasmid

目的克隆人PLAGL2基因片段并插入(TetO)7-CMV表达载体。方法采用PCR技术扩增出人PLAGL 2基因的两个DNA片段,经酶连后获得约2kb的目的DNA片段并插入(TetO)7-CMV载体中,PCR筛选阳性转化株并分析重组质粒的DNA序列。结果DNA测序结果显示重组质粒(TetO)7-CMV-PLAGL 2序列正确。结论扩增并连接了人PLAGL2基因的两个DNA片段,最终成功构建了人PLAGL2基因的表达载体质粒—(TetO)7-CMV-PLAGL2。

Objective： To clone DNA fragment of human PLAGL 2 gene and insert it into （TetO）7 - CMV expressive vector. Methods： The two DNA fragment of human PLAGL 2 gene was amplified from human genomic DNA respectively, then combined them with T4 DNA ligase, and then the 2kb combined DNA fragment was subcloned into （TetO）7 -CMV expressive vector. The plasmid was determined by DNA sequencing. Result： The results of DNA sequencing showed that the sequence of recombined DNA plasmid was correct. Conclusion： The DNA fragment of human PLAGL 2 gene was amplified and combined correctly, and recombined DNA plasmid of （TetO）7-CMV- PLAGL 2 was constructed successfully.