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果蝇SMART检测中紫外线刺激重组的分析。

Analysis of UV-stimulated recombination in the Drosophila SMART assay.

作者信息

Kaya Bülent, Kocaoğlu Serap, Demir Eşref

机构信息

Department of Biology, Faculty of Arts and Sciences, Akdeniz University, Antalya, Turkey.

出版信息

Environ Mol Mutagen. 2006 Jun;47(5):357-61. doi: 10.1002/em.20215.

Abstract

The UV component of solar radiation is classified into UVA (320-400 nm), UVB (290-320 nm), and UVC (200-290 nm). Although all three types of UV light are capable of damaging biological systems, the earth's atmosphere filters out UVC, and a portion of UVB. In this study, we evaluated the induction of mutation and recombination by different wavelengths of UV light in the wing spot test of Drosophila melanogaster (Somatic Mutation and Recombination Test, SMART). Third-instar larvae that were trans-heterozygous for the third chromosome recessive markers, multiple wing hairs (mwh) and flare-3 (flr(3)), were exposed to different doses of UVA (at 365 nm), UVB (at 312 nm) or UVC (at 254 nm), and transferred to standard Drosophila culture medium. Feeding ended with pupation of the surviving larvae, and the genetic changes induced in the somatic cells of the wing's imaginal discs lead to the formation of mutant clones on the wing blade. Point mutation, chromosome breakage, and mitotic recombination produce single spots, while twin spots are produced only by mitotic recombination. Exposure to 500-4,000 J/cm(2) UVA did not increase the frequency of mutant spots. UVB doses of 200, 250, 300, 350, and 400 J/cm(2) increased the frequency of all categories of spots, indicating that UVB was potentially both mutagenic and recombinogenic. Assays run in balancer-heterozygous flies (which are insensitive to recombination) indicated that the fraction of mutants in trans-heterozygous flies due to recombination increased from 48.57% at 200 J/cm(2) UVB to 98.30% at 400 J/cm(2) UVB. While 140-480 J/cm(2) of UVC was not genotoxic, UVC produced a strong toxic response at doses higher than 140 J/cm(2). The results of this study indicate that UVB was much more active than UVC or UVA in the SMART assay, and that UVB was highly recombinogenic.

摘要

太阳辐射的紫外线成分可分为UVA(320 - 400纳米)、UVB(290 - 320纳米)和UVC(200 - 290纳米)。虽然这三种紫外线都能够损害生物系统,但地球大气层会滤除UVC以及一部分UVB。在本研究中,我们在黑腹果蝇的翅斑试验(体细胞突变和重组试验,SMART)中评估了不同波长紫外线诱导的突变和重组情况。对第三染色体隐性标记多翅毛(mwh)和flare - 3(flr(3))为反式杂合的三龄幼虫,暴露于不同剂量的UVA(365纳米)、UVB(312纳米)或UVC(254纳米),然后转移到标准果蝇培养基中。饲养至存活幼虫化蛹结束,翅成虫盘体细胞中诱导的基因变化会导致翅片上形成突变克隆。点突变、染色体断裂和有丝分裂重组产生单斑,而双斑仅由有丝分裂重组产生。暴露于500 - 4000 J/cm²的UVA并未增加突变斑的频率。200、250、300、350和400 J/cm²的UVB剂量增加了所有类型斑的频率,表明UVB可能兼具致突变性和重组性。在平衡子杂合果蝇(对重组不敏感)中进行的试验表明,反式杂合果蝇中由于重组产生的突变体比例从200 J/cm² UVB时的48.57%增加到400 J/cm² UVB时的98.30%。虽然140 - 480 J/cm²的UVC没有遗传毒性,但UVC在高于140 J/cm²的剂量下会产生强烈的毒性反应。本研究结果表明,在SMART试验中UVB比UVC或UVA更具活性,且UVB具有高度重组性。

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