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目的克隆人p27kiplcDNA并将其转染至Tca8113细胞中,建立稳定转染p27kipl基因的细胞株。方法用PT-PCR法获得人p27kiplcDNA,将其克隆人pcDNA3载体上,并将重组表达质粒经脂质体介导转染Tca8113细胞,经CA18抗性筛选后用PCR鉴定。结果克隆出人的p27kiplcDNA并将其转染至Tca8113细胞中,获得了稳定转染p27kipl基因的细胞株。结论用分子生物学技术克隆了人p27kipl。基因,获得稳定转染p27kipl基因的细胞株,为进一步研究p27kip基因治疗舌癌的实验研究奠定基础。
Objective To clone human p27kiplcDNA and transfect it into Tca8113 cells to establish a cell line stably transfected with p27kipl gene. Methods Human p27kiplcDNA was obtained by PT-PCR and cloned into human pcDNA3 vector. The recombinant plasmid was transfected into Tca8113 cells by lipofectamine 2000 and identified by PCR with CA18. Results Human p27kiplcDNA was cloned and transfected into Tca8113 cells. The cell line stably transfected with p27kipl gene was obtained. Conclusion Human p27kipl was cloned by molecular biology technique. Gene to obtain stably transfected p27kipl gene cell strain, which lays the foundation for further study of p27kip gene therapy for tongue cancer.