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| 甲基硝基亚硝基胍诱发的遗传不稳定vero细胞DNA体外复制的保真度 | |
| 引用本文： | 冯朝晖,余应年,陈星若. 甲基硝基亚硝基胍诱发的遗传不稳定vero细胞DNA体外复制的保真度[J]. 中国药理学与毒理学杂志, 1998, 12(2): 144-148 |
| 作者姓名： | 冯朝晖 余应年 陈星若 |
| 作者单位： | 浙江医科大学病理生理学教研室及分子生物学实验室 |
| 摘 要： | 利用在猴肾vero细胞抽提物中建立的DNA体外复制系统，对穿梭质粒pZ189 DNA进行有效的复制后，将复制产物转化至E.coli MBM7070，观察pZ189质粒DNA中突变靶基因supF tRNA基因突变的情况．在烷化剂甲基硝基亚硝基胍(N-methyl-N′-nitro-N-nitrosoguanidine)诱发的遗传不稳定(genetic instability) vero细胞胞浆抽提物中进行pZ189质粒DNA复制, 发现经过体外复制的pZ189质粒中supF tRNA基因突变率高出对照组5-8倍(P<0.05)，证实该细胞DNA复制系统的复制保真度明显降低. |
| 关 键 词： | 甲基硝基亚硝基胍； 诱变； 质粒； DNA复制； 细胞 vero |
| 收稿时间： | 1997-06-02 |
Fidelity of DNA replication in vitro by extracts of genetically instable vero cells induced by N-methyl-N′-nitro-N-nitrosoguanidine |
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| FENG Zhao-Hui, YU Ying-Nian, CHEN Xing-Ruo. Fidelity of DNA replication in vitro by extracts of genetically instable vero cells induced by N-methyl-N′-nitro-N-nitrosoguanidine[J]. Chinese Journal of Pharmacology and Toxicology, 1998, 12(2): 144-148 | |
| Authors: | FENG Zhao-Hui YU Ying-Nian CHEN Xing-Ruo |
| Affiliation: | (Department of Pathophysiology and Laboratory of Medical Molecular Biology, Zhejiang Medical University, Hangzhou 310031) |
| Abstract: | Shuttle plasmid pZ189 DNA was replicated efficiently by the in vitro DNA replication system derived from extracts of monkey kidney vero cells exhibiting genetic instability which was induced by exposure to 0.2 μmol·L-1 N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) for 2.5 h. The replicated pZ189 DNA was used to transform E.coli MBM7070 to examine the nontargeted mutation occurring in the mutagenesis target, supF tRNA gene, in pZ189 DNA. It was identified that the mutation frequency of supF tRNA gene in pZ189 replicated in the above extracts was 5 to 8 times higher than that replicated in extracts of untreated vero cells (P<0.05). The result demonstrated that the fidelity of DNA replication in vitro by extracts of MNNG pretreated vero cells decreases obviously, which may result from the functional defects in DNA replication system, and play a significant role in the genesis of MNNG-induced genetic instability in vero cells. |
| Keywords: | N-methyl-N′-nitro-N-nitrosoguanidine mutagenesis plasmids DNA replication cells vero |
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