分泌高滴度重组人糖皮质激素β受体正义和反义cDNA逆转录病毒细胞系的建立

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G418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G418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2×105 CFU/ml和1.8×105 CFU/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。

Construction of cell lines producing high-titer recombinant retroviruses of message and antisense cDNA of human glucocorticoid receptor β

OBJECTIVE: To construct cell lines producing high-titer recombinant retroviruses of human glucocorticoid receptor beta (hGRbeta). METHODS: The recombinant retrorinal vectors of message and antisense cDNA of hGRbeta were transfected into PA317 cells by liposomes, respectively, and cell colonies were selected with G418. The titers of viruses from the 2 cell culture supernatants were determined by colony numbers produced in NIH3T3 cells. RESULTS: The presence of aimed RNAs in supernatant of the G418-resistant cell cultures were detected by RT-PCR and subsequent sequence analysis. The titers of retroviruses were 2. 0 x 10(5) and 1.8 x 10(5), each for recombinant message and antisense hGRbeta cDNA. CONCLUSION: We demonstrated here the obtaining of transfect PA317 cells that produce recombinant retroviruses containing message and antisense RNAs of hGRbeta with highly infective efficiencies.