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目的构建TPT1真核报告表达载体pEGFPN3TPT1,为进一步研究其功能奠定基础。方法通过PCR和酶切的方法获取具有黏性末端的TPT1基因的全长开放读码框,将其定向连入真核表达载体pEGFPN3绿色荧光报告基因的氨基侧,构成pEGFPN3TPT1的真核报告表达载体,通过酶切和测序鉴定TPT1的正确插入。结果酶切和测序均证实TPT1正确插入了真核报告表达载体pEGFPN3TPT1中。结论成功构建了TPT1的真核报告表达载体pEGFPN3TPT1。
Objective To construct TPT1 eukaryotic expression vector pEGFPN3TPT1, which laid the foundation for further study of its function. Methods The open reading frame (ORF) of TPT1 gene with cohesive ends was obtained by PCR and digestion. The full-length open reading frame (ORF) of TPT1 gene was cloned into the amino-terminal of the green fluorescent reporter gene pEGFPN3. The eukaryotic expression vector pEGFPN3TPT1 , Correct insertion of TPT1 was identified by restriction and sequencing. Results Both digestion and sequencing confirmed that TPT1 was correctly inserted into the eukaryotic expression vector pEGFPN3TPT1. Conclusion The eukaryotic expression vector pEGFPN3TPT1 of TPT1 was successfully constructed.