转基因果蝇DNA聚合酶基因过表达对DNA修复和重组的影响。

Idengaku zasshi Pub Date : 1994-02-01 DOI:10.1266/jjg.69.21
M A Yoo, W H Lee, H Y Ha, J R Ryu, M Yamaguchi, K Fujikawa, A Matsukage, S Kondo, Y Nishida
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引用次数: 0

摘要

将大鼠DNA聚合酶β (pol β) cDNA融合到果蝇肌动蛋白5C基因(简称pol β)的增强子-启动子区和多a信号序列转移到果蝇基因组中。研究了在不同染色体上具有转基因pol β基因的4个转基因菌株中的3个。在转基因菌株的胚胎和转染了携带pol - β载体的培养细胞中,pol - β转录物水平和聚合活性水平均显著升高。普遍认为DNA聚合酶β参与DNA修复的观点并不支持,因为观察到一对正常菌株和一对pol β菌株,以及另一对DNA修复缺陷双突变菌株mei-9 mei-41 (DNA修复缺陷双突变)菌株和pol β mei-9 mei-41菌株在紫外线照射后,每对菌株的存活率没有差异。DNA聚合酶β参与重组的另一个观点得到了研究结果的支持,即转基因pol β菌株的自发重组频率,无论是减数分裂还是有丝分裂,都明显高于非转基因菌株。然而,pol β菌株中增强的重组频率可能反映了过量生产的pol β蛋白对染色体稳定性的间接影响。无论大鼠pol - β的直接作用是什么,转基因pol - β果蝇将有助于研究pol - β的生理作用和重组机制。
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Effects of DNA polymerase beta gene over-expressed in transgenic Drosophila on DNA repair and recombination.

DNA polymerase beta (pol beta) cDNA of rat fused to an enhancer-promoter region plus a poly(A) signal sequence of actin 5C gene of Drosophila (abbreviated pol beta) was transferred to the Drosophila genome. Three of four constructed transgenic strains possessing transgene pol beta on different chromosomes were studied. Levels of the pol beta transcript and those of the polymerization activity of pol beta were markedly elevated in cultured cells transfected with pol beta-bearing vectors as well as in embryos of the transgenic strains. The popular idea that DNA polymerase beta participates in DNA repair was not supported by the observation that a pair of a normal and a pol beta strain, and the other pair of a mei-9 mei-41 (DNA-repair deficient double mutations) strain and a pol beta mei-9 mei-41 strain, showed no difference in survival within each pair after treatment with ultraviolet light, methylmethane sulfonate and mitomycin C. The other idea that DNA polymerase beta participates in recombination was supported by the findings that spontaneous frequency of recombination, either meiotic or mitotic, is significantly higher in a transgenic pol beta strain than in a non-transgenic strain. The enhanced recombination frequency in the pol beta strain may, however, reflect an indirect effect of over-produced pol beta proteins on chromosomal stability. Whatever the direct effect of rat pol beta is, the transgenic pol beta flies will be useful for study of the physiological role of pol beta and the mechanism of recombination.

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