The mouse lymphoma assay detects recombination, deletion, and aneuploidy.
The mouse lymphoma assay detects recombination, deletion, and aneuploidy.
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小鼠淋巴瘤检测可检测重组、缺失和非整倍性。
DOI：
10.1093/toxsci/kfp037
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
M. Moore
中科院分区：
文献类型：
--
作者：
Jianyong Wang;J. Sawyer;Ling Chen;Tao Chen;M. Honma;N. Mei;M. Moore
小鼠淋巴瘤试验（MLA）使用L5178Y/Tk(+/-)-3.7.2C小鼠淋巴瘤细胞系的胸苷激酶(Tk)基因作为报告基因来评估化学和物理制剂的致突变性。 MLA 被美国食品和药物管理局和美国环境保护局推荐为遗传毒理学筛查的首选体外哺乳动物细胞突变检测，因为它可以检测广泛的遗传改变，包括点突变和染色体突变。然而，MLA 可检测的染色体突变的具体类型需要进一步澄清。为此，使用三种化学物质（包括两种断裂剂和一种aneugen（3'-叠氮基-3'-脱氧胸苷、丝裂霉素C 和紫杉醇））来诱导Tk 突变体。杂合性丢失（LOH）分析用于选择突变体，这些突变体可以提供关于它们是否由缺失、有丝分裂重组或非整倍性产生的信息。然后结合其他方法、G 带分析、染色体涂色和检测 Tk 基因拷贝数 (CN) 的实时 PCR 方法来提供详细分析。 LOH 涉及至少 25% 的 11 号染色体、正常核型和 Tk CN 为 2，表明突变体是重组产生的；而 LOH 与核型上可见的 11 号染色体缺失和 Tk CN 为 1 相结合表明是缺失。对于在 11 号染色体上的每个微卫星标记处显示 LOH 的突变体，使用 G 带结合结合染色体涂色分析证实了非整倍性。从该分析中可以清楚地看出，小鼠淋巴瘤 Tk 突变体可能是由重组、缺失和非整倍性引起的。
The mouse lymphoma assay (MLA) uses the thymidine kinase (Tk) gene of the L5178Y/Tk(+/-)-3.7.2C mouse lymphoma cell line as a reporter gene to evaluate the mutagenicity of chemical and physical agents. The MLA is recommended by both the United States Food and Drug Administration and the United States Environmental Protection Agency as the preferred in vitro mammalian cell mutation assay for genetic toxicology screening because it detects a wide range of genetic alterations, including both point mutations and chromosomal mutations. However, the specific types of chromosomal mutations that can be detected by the MLA need further clarification. For this purpose, three chemicals, including two clastogens and an aneugen (3'-azido-3'-deoxythymidine, mitomycin C, and taxol), were used to induce Tk mutants. Loss of heterozygosity (LOH) analysis was used to select mutants that could be informative as to whether they resulted from deletion, mitotic recombination, or aneuploidy. A combination of additional methods, G-banding analysis, chromosome painting, and a real-time PCR method to detect the copy number (CN) of the Tk gene was then used to provide a detailed analysis. LOH involving at least 25% of chromosome 11, a normal karyotype, and a Tk CN of 2 would indicate that the mutant resulted from recombination, whereas LOH combined with a karyotypically visible deletion of chromosome 11 and a Tk CN of 1 would indicate a deletion. Aneuploidy was confirmed using G-banding combined with chromosome painting analysis for mutants showing LOH at every microsatellite marker on chromosome 11. From this analysis, it is clear that mouse lymphoma Tk mutants can result from recombination, deletion, and aneuploidy.
通过杂合性缺失 (LOH) 和整个 11 号染色体染色分析大和小菌落 L5178Y tk-/- 小鼠淋巴瘤突变体：重组检测。
影响因子：
2.7
作者：
Liechty,MC;Scalzi,JM;Sims,KR;CrosbyJr,H;Spencer,DL;Davis,LM;Caspary,WJ;Hozier,JC
通讯作者：
Hozier,JC