【摘要】 本文通过用已知诱变剂EMS对基于EB₂病毒的穿梭载体pMCi5在小鼠3T3细胞中的诱变作用的研究,初步建立了一种能在DNA水平研究哺乳动物细胞诱变作用的快速实验系统。pMCi5质粒带有细菌lacI基因,可作为体细胞诱变研究的靶基因,最后通过转化大肠杆菌,在含X-gal的培养基中快速检测lacI突变。该系统的自发突变率小于1.21×10^-4,300μ/mlEMS的诱发突变率为3.8×10^-3。经酶谱分析得到的7个突变子,在DNA水平上没有大的改变,EcoRl酶切位点上无点突变发生。更多还原

【Abstract】 A series of experimental researches was reported involved in developing a short-time mutagenesis system to detect gene mutation in mammalian cells at DNA level by using shuttle vector. In this system the bacterial lacI gene carried by shuttle vector pMCi5 used as a target for mutagenesis in mouse cell. An EB virus based plasmid pMCi5 was transformed into mouse 3T3 cell and exposed to chemical mutagen ethyl methanesulfonte (EMS). Later returned the treated plasmid to E. coli strain MC1061F’ 150kan and plate the transformers on the special medium containing X-gal for rapid detection and analysis of lacI mutation.The spontaneous mutant frequency of this system is less than 1.21X10-4. After treated by 300ug/ml EMS, induced mutant frequency increased 3.8X10-3. Neither has gross alteration been discovered for 7 lacI mutants at DNA level, nor has point mutation of EcoRI site been found yet by analysis with restriction enzyme EcoRI.更多还原