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  

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

作　　者：朱敏[1] 何庆南[2] 周钢[1] 张磊[2] C.I.Msangi 胡维新[1] 

机构地区：[1]生物科学与技术学院分子生物学研究中心,长沙410078 [2]湘雅二医院儿科及小儿肾病学实验室,长沙410011

出　　处：《中南大学学报（医学版）》2004年第5期540-543,共4页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金 (30 0 0 0 1 84;3990 0 1 59)

摘　　要：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 。Objective To construct cell lines producing high-titer recombinant retroviruses of human glucocorticoid receptor β(hGRβ). Methods The recombinant retrorinal vectors of message and antisense cDNA of hGRβ 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 2.0×10 5 and 1.8×10 5,each for recombinant message and antisense hGRβ cDNA. Conclusion We demonstrated here the obtaining of transfect PA317 cells that produce recombinant retroviruses containing message and antisense RNAs of hGRβ with highly infective efficiencies.

关 键 词：反义CDNA 逆转录病毒 Β受体 糖皮质激素 分泌 细胞系 高滴度 细胞克隆 全长cDNA 质粒 

分 类 号：R346[医药卫生—基础医学]