pS189／FLCⅢ—2诱变检测系统的研究  

THE ESTABLISHMENT OF pS189/FLCⅢ 2 SYSTEM FOR MUTAGENESIS STUDY

作　　者：易绍萱[1,2,3] 李大成[1,2,3] 周晓柳 王浩[1,2,3] 傅继梁[1,2,3] 

机构地区：[1]第三军医大学分子生物学教研室 [2]华西医科大学分子生物学教研室 [3]第二军医大学生物学教研室

出　　处：《癌变．畸变．突变》1996年第5期276-282,共7页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家八五攻关项目;美国CMB基金

摘　　要：本文报道优选基于ＳＶ４０的短暂复制型穿梭质粒ｐＳ１８９和新生儿羊膜上皮细胞ＦＬＣⅢ—２亚克隆，组成穿梭质粒／哺乳动物细胞诱变检测系统。该系统中ＳｕｐＦ的自发突变型频率仅为１．７×１０－５，低于已报道的其它短暂复制型质粒和该质粒在其它细胞中复制时ＳｕｐＦ基因的自发突变率。各种剂量的ＭＮＮＧ的诱发突变频率随诱变剂剂量的增加而升高。ＭＮＮＧ对ＦＬ细胞毒性较大，仅０．３２μｇ／ｍｌ就可使ＦＬ几乎完全失去生长分裂形成克隆的能力，但在短时期内对依赖于病毒复制起点的ＤＮＡ复制效率影响不大，突变频率却显著升高。突变质粒的限制性内切酶分析和靶基因的ＰＣＲ分析等表明，检出突变大多数为点突变或微小缺失，该系统具有方便快速检出点突变的能力，可成为检测诱变剂、抗诱变剂作用强度和特性的分子检测系统。A mutagenesis experimental system has been established for detecting mutagens and elucidation the mechanism of mutagenesis of DNA level in mammalian cells,using a SV40 based transient replication shuttle vector plasmid (pS189) and a human amnion cell strain(FL).The mutant frequency of control in this system is only 1.7×10 -5 .After treated with 0.32μg/ml MNNG,the induced mutant frequency increased significantly(P<0.05).The mutants were analysed by the agrose gel electrophoresis,restriction enzyme analysis and PCR amplication of SupF gene.It is indicated that the majority of the SupF mutants induced by MNNG or EMS in the pS189/FLCⅢ 2 system may be point mutations.

关 键 词：穿梭质粒 诱发突变 pS189 环境致癌 检测系统 

分 类 号：R994.6[医药卫生—毒理学]