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目的 :建立分泌重组人糖皮质激素β受体cDNA的逆转录病毒的细胞系。方法 :脂质体介导含人糖皮质激素β受体正义和反义全长cDNA的重组逆病毒质粒分别转染包装细胞PA317,经G4 18筛选抗性克隆。结果 :细胞培养上清液的RT PCR联合序列分析表明G4 18抗性PA317细胞克隆能稳定合成并分泌相应的重组病毒颗粒 ;NIH3T3细胞测定的正、反义cDNA重组病毒滴度分别为 2× 10 5CFU/ml和 1.8× 10 5CFU/ml。结论 :成功建立了能分别产生重组hGRβ正义和反义cDNA逆转录病毒的高滴度细胞系 ,为后续研究工作打下坚实基础
OBJECTIVE: To establish a retrovirus cell line that secretes recombinant human glucocorticoid receptor β cDNA. METHODS: Lipofectamine mediated recombinant retroviral plasmid containing sense and antisense full length cDNA of human glucocorticoid receptor were transfected into PA317 packaging cells, and the resistant clones were screened by G418. Results: RT-PCR analysis of cell culture supernatants showed that G418-resistant PA317 cells could stably synthesize and secrete the corresponding recombinant virus particles. The titer of the recombinant antisense cDNA of NIH3T3 cells was 2 × 10 5 CFU / ml and 1.8 × 10 5 CFU / ml. CONCLUSION: High-titer cell lines capable of producing recombinant hGRβ sense and antisense cDNA retroviruses were successfully established, laying a solid foundation for subsequent research