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目的:构建并鉴定含有plap-1基因和已知FLAG序列的真核细胞表达载体pBABE-hygro-PLAP-1。方法:采用PCR扩增PLAP-1的目的片段,然后连接到已知FLAG序列的载体p3×FLAG-CMV-10上;再次通过PCR扩增出PLAP-1的目的片段及已知序列的FLAG,最后将其插入真核细胞表达载体pBABE-hygro的多克隆位点上。对该重组体进行酶切鉴定和测序验证。结果:通过对重组质粒进行酶切鉴定以及DNA序列测定分析,证明真核表达载体pBABE-hygro-PLAP-1构建成功。结论:成功构建plap-1基因的真核表达载体,为进一步研究plap-1基因的功能奠定了实验基础。
Objective: To construct and identify eukaryotic expression vector pBABE-hygro-PLAP-1 containing plap-1 gene and known FLAG sequence. Methods: The target fragment of PLAP-1 was amplified by PCR and ligated into vector p3 × FLAG-CMV-10 with known FLAG sequence. The target fragment of PLAP-1 and the known sequence of FLAG were amplified by PCR. Finally, it was inserted into the eukaryotic cell expression vector pBABE-hygro multiple cloning site. The recombinants were identified by restriction enzyme digestion and sequencing. Results: The eukaryotic expression vector pBABE-hygro-PLAP-1 was successfully constructed by restriction enzyme digestion and DNA sequence analysis. Conclusion: The eukaryotic expression vector of plap-1 gene was successfully constructed, which laid the experimental foundation for further study on the function of plap-1 gene.