STUDY ON THE MUTATION SPECTRUM OF hprt GENE IN HeLa MR CELL INDUCED BY γ-RAY

Zhao Yanhua; Xing Ruiyun; Ma Ping; Wang Huichen; Xia Shouxuan

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STUDY ON THE MUTATION SPECTRUM OF hprt GENE IN HeLa MR CELL INDUCED BY γ-RAY

更新时间：2023-04-18

- STUDY ON THE MUTATION SPECTRUM OF hprt GENE IN HeLa MR CELL INDUCED BY γ-RAY
- Journal of Radiation Research and Radiation Processing Vol. 16, Issue 1, Pages: 20-24(1998)
- 作者机构：
  
  1.北京放射医学研究所　北京　100850
- 作者简介：
- 基金信息：
  
  THe project was supported by The National Science Foundation of China(39130050)
- DOI：
  CLC：
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Zhao Yanhua, Xing Ruiyun, Ma Ping, et al. STUDY ON THE MUTATION SPECTRUM OF hprt GENE IN HeLa MR CELL INDUCED BY γ-RAY. [J]. Journal of Radiation Research and Radiation Processing 16(1):20-24(1998)
DOI：

Zhao Yanhua, Xing Ruiyun, Ma Ping, et al. STUDY ON THE MUTATION SPECTRUM OF hprt GENE IN HeLa MR CELL INDUCED BY γ-RAY. [J]. Journal of Radiation Research and Radiation Processing 16(1):20-24(1998) DOI：

摘要

将正常HeLa MR细胞用,60,Co γ射线分别进行0～8.0Gy照射后，在含6-巯基鸟嘌呤（6-TG）的培养基中克隆和筛选hprt基因突变细胞。细胞的突变频率与,γ,射线的照射剂量有关，共获得3个自然突变的细胞克隆和18个γ射线诱导的突变细胞克隆，用8对hprt寡核苷酸引物进行多聚酶链式反应（PCR）,从培养细胞提取DNA中分别扩增hprt基因的9个外显子片段，分析突变细胞的hprt基因外显子突变情况。结果表明，在辐射致突的克隆中，约77.8%（14/18）的hprt基因突变为外显子缺失，其中2个突变是外显子全部缺失，而12个是外显子部分缺失。另外，约有22.2%（4/18）的突变无明显的PCR扩增变化，属于点突变，且突变程度与照射剂量有关。在3个自然突变克隆中，点突变和缺失突变的比例分别为67%（2/3）和33%（1/2）,突变程度远远低于照射组。这对于估计辐射剂量和辐射远后效应观察，揭示辐射致突的分子机理提供了信息。

Abstract

Normal HeLa MR cells were irradiated by ,60,Co γ-ray in the dose range of 0~8Gy and the mutants of hprt gene were cloned in the 6-TG containing selective culture and assayed. Three natural spontaneous mutants and eighteen γ-ray induced hprt mutants were recovered. The mutation spectra of hprt exons in all of them were analyzed by polymerase chain reaction (PCR) using 8 pairs of hprt oligonucleotide primers to amplify hprt exons from cellular DNA extracts. The result showed that in the mutants induced by γ-ray, about 77.8% (14/18) of the hprt mutations were deleted,among which 2 were total deletion, the other 12 were partial deletion,and 22.2% (4/18) ,which did not show noticeable changes in the PCR pattern,were point mutation. The mutation degree of hprt exon deletion was found to be dependent on the irradiation dose. However,about 67% (2/3) point mutants and 33% (1/3) deleted mutants were showed in the natural mutants. Their mutation degree was far lower than that of the γ-ray induced mutants. These r esults might be beneficial for further study of the molecular mechanism of mutation induced by γ-ray irradiation as well as for the estimation of irradiation dose.

关键词

γ辐射hprt基因基因突变谱

Keywords

γ-irradiationhprt geneGene mutation spectrum

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