癌变·畸变·突变 ›› 2005, Vol. 17 ›› Issue (5): 261-264.doi: 10.3969/j.issn.1004-616x.2005.05.002

• 论著 • 上一篇    下一篇

小鼠淋巴瘤L5178Y细胞tk基因自发分子突变研究

袁 健;刘胜学;刘晋炜;曹 佳   

  1. 第三军医大学军事预防医学院卫生毒理学教研室,重庆 400038
  • 收稿日期:2004-09-16 修回日期:2005-01-17 出版日期:2005-09-30 发布日期:2005-09-30

The Study of Spontaneous Mutations at the tk Gene of L5178Y Mouse Lymphoma Cells

YUAN Jian;LIU Sheng-xue; LIU Jin-yi ; CAO Jia   

  1. Department of Hygiene Toxicology, College of Military Preventive Medicine, Third Military Medical University, Chongqing 400038, China
  • Received:2004-09-16 Revised:2005-01-17 Online:2005-09-30 Published:2005-09-30

摘要: 背景与目的: 研究小鼠淋巴瘤L5178Y3.2.7c-tk +/-细胞tk基因的自发突变频率和分子突变。 材料与方法: 进行4次平行试验,计算自发突变频率。选择tk基因自发突变子(tk-/-),提取基因组DNA, 经等位基因特异性PCR扩增,杂合性缺失(Loss of heterozygosity,LOH)分析和测序等技术,分析其分子突变谱。 结果: L5178Y细胞tk基因的平均自发突变频率为145×10-6,tk基因自发突变LOH平均发生率为79.8%。由LOH分析的试验结果显示,自发突变的LOH在大集落(Large clone,LC)与小集落(Small clone,SC)之间差异无统计学意义。序列分析证实,自发tk基因点突变主要为碱基置换。 结论: tk基因的自发突变是以功能性等位基因缺失为主的分子突变,其中点突变以碱基置换为主。

关键词: 自发突变, tk位点, 杂合性缺失, 分子突变

Abstract: BACKGROUND & AIM: To investigate the mutation frequency and molecular mutation of spontaneous mutations on the tk gene of mouse lymphoma cells. MATERTAL AND METHODS:Performing four times parallel test. Mutation frequency of tk locus were analyzed by the mouse lymphoma assay(microitier procedure), and Loss of heterozygosity(LOH) analysis withallic special PCR. Subsequently, we determined the DNA sequence of negative LOH's clones. RESULTS:Loss of heterozygosity analysis of the tk locus revealed that the most spontaneous mutants lost the functional tk allele (average LOH 79.8%). LOH of spontaneous mutations showed there were no significant difference between large clone(LC) and small clone(SC).Spontaneous mutations were substitutions, and most of them were transversions. CONCLUSION: Functional tk allic gene lose was major mutational event in spontaneous mutations and point mutations were substitutions.

Key words: spontaneous mutations, tk locus, loss of heterozygosity, molecular mutation