MyD88干涉慢病毒颗粒的包装和鉴定  

Packaging and identification of lentivirus granules interfering MyD88

作　　者：周仕香[1,2] 王治东[2] 董波[2] 张学清[2] 韩春光[2] 高荣莲[2] 刘芬菊[1] 陈肖华[2] 

机构地区：[1]苏州大学医学部放射医学与公共卫生学院,苏州215123 [2]军事医学科学院放射与辐射医学研究所,北京100850

出　　处：《军事医学科学院院刊》2010年第2期150-152,共3页Bulletin of the Academy of Military Medical Sciences

基　　金：北京市自然科学基金(7102121)

摘　　要：目的制备MyD88分子的慢病毒干涉颗粒,建立稳定下调MyD88的HEK293细胞系。方法将携带特异性干涉序列的DNA片段克隆入干涉质粒pSicoR中,并制备携带不同干涉序列的慢病毒颗粒。将慢病毒颗粒感染HEK293细胞,建立稳定细胞系,经RT-PCR和报告基因分析方法确定不同干涉序列对MyD88表达的下调及信号通路激活的阻断作用。结果成功制备携带干涉序列的慢病毒颗粒,并获得稳定感染慢病毒的HEK293细胞系,经鉴定发现感染912位序列的细胞MyD88mRNA表达水平下调为对照组的28%,对CBLB502的激活作用下降为对照组的24%。结论成功制备MyD88的慢病毒干涉颗粒,可用于建立MyD88稳定下调的细胞系。Objective To prepare the lentivirus granules interfering MyD88 and establish HEK293 cell line consistently down-regulating MyD88.Methods The lentivirus granules bearing various interfering sequences were prepared by cloning DNA segments carrying specific interfering sequences into interfering plasmid pSicoR.The stable cell lines were established after infecting HEK293 cells with lentivirus granules.RT-PCR and reporter gene analysis were employed to determine the inhibitory effects of various interfering sequences on MyD88 expression and signal pathway activation.Results The lentivirus granules bearing various interfering sequences were successfully prepared.The stable HEK293 cell lines infected by lentivirus were obtained.MyD88 mRNA expression level from 912 sequence infected cells was down-regulated to 28% of that of normal control.CBLB502 activation was down-regulated to 24% of that of normal control.Conclusion Successful preparation of lentivirus granules interfering MyD88 can facilitate establishment of MyD88 stably down-regulated cell lines.

关 键 词：髓样分化相关蛋白 慢病毒 RNA干涉 细胞系 质粒 

分 类 号：R73-3[医药卫生—肿瘤]