ANALYSIS OF MUTATION IN HUMAN-CELLS BY USING AN EPSTEIN-BARR-VIRUS SHUTTLE SYSTEM
ANALYSIS OF MUTATION IN HUMAN-CELLS BY USING AN EPSTEIN-BARR-VIRUS SHUTTLE SYSTEM
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DOI：
10.1128/mcb.7.1.379
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发表时间：
1987-01-01
影响因子：
5.3
通讯作者：
CALOS, MP
中科院分区：
文献类型：
--
作者：
DUBRIDGE, RB;TANG, P;CALOS, MP
我们开发了高度敏感的穿梭载体系统，用于检测使用自主复制的EB病毒（EBV）衍生物在人类细胞中形成的突变。携带细菌lacI基因作为突变靶的EBV载体在人细胞中建立，随后返回大肠杆菌用于lacI突变的快速检测和分析。通过建立lacI-EBV载体产生的大多数克隆细胞系显示自发的LacI-频率小于10-5，并且适合于诱导突变的研究。分离克隆系的能力代表了EBV载体在突变研究中优于瞬时复制穿梭载体（如来源于猿猴病毒40的那些）的主要优势。DNA序列的变化，确定了61 lacI突变的细胞系暴露于N-亚硝基-N-甲基脲诱导。34个lacI无义突变中的33个和27个错义突变中的26个涉及G. cntdot。C到A. cntdot. T转换。这些数据为癌症的突变理论提供了支持。
We developed highly sensitive shuttle vector systems for detection of mutations formed in human cells using autonomously replicating derivatives of Epstein-Barr virus (EBV). EBV vectors carrying the bacterial lacI gene as the target for mutation were established in human cells and later returned to Escherichia coli for rapid detection and analysis of lacI mutations. The majority of the clonal cell lines created by establishment of the lacI-EBV vector show spontaneous LacI- frequencies of less than 10-5 and the are suitable for studies of induced mutation. The ability to isolate clonal lines represents a major advantage of the EBV vectors over transiently replicating shuttle vectors (such as those derived from simian virus 40) for the study of mutation. The DNA sequence changes were determined for 61 lacI mutations induced by exposure of one of the cell lines to N-nitroso-N-methylurea. A total of 33 of 34 lacI nonsense mutations and 26 of 27 missense mutations involve G .cntdot. C to A .cntdot. T transitions. These data provide support for the mutational theory of cancer.