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目的 :克隆小鼠 4 1BBL基因 ,构建其真核表达载体 ,并观察其在哺乳动物细胞中的表达 .方法 :取C5 7BL/ 6小鼠脾细胞 ,经PHA诱导后 ,以RT PCR克隆 4 1BBLcDNA ,测序 ,构建 pCDNA3.1 (+) m4 1BBL真核表达质粒 ,转染COS 7细胞 ,RT PCR检测m4 1BBLmRNA表达 ,间接免疫荧光法和流式细胞仪检测 4 1BBL蛋白的表达 .结果 :从小鼠脾细胞中克隆到m4 1BBLcDNA ,经测序完全正确 ,所构建的m4 1BBL质粒在COS 7细胞中获得高效表达 .结论 :小鼠 4 1BBLcDNA基因克隆、真核表达载体构建及其在COS 7中的表达均获成功 ,为进一步研究其功能奠定了基础
OBJECTIVE: To clone 4 1BBL gene of mouse and construct its eukaryotic expression vector and observe its expression in mammalian cells.Methods: After spleen cells from C5 7BL / 6 mice were induced by PHA, 4 1BBL cDNA was cloned by RT PCR , And the eukaryotic expression plasmid pCDNA3.1 (+) m4 1BBL was constructed and transfected into COS-7 cells.The mB 1BBL mRNA expression was detected by RT-PCR and the expression of 4 1BBL protein was detected by indirect immunofluorescence and flow cytometry.Results: The m4 1BBL cDNA was cloned from spleen cells and sequenced completely.The constructed m4 1BBL plasmid was highly expressed in COS7 cells.Conclusion: The cloning and eukaryotic expression vector of mouse 4 1BBL cDNA and its expression in COS 7 All were successful, laying the foundation for further research on their functions