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人体睾丸中体细胞DNA修复基因的表达。

Expression of somatic DNA repair genes in human testes.

作者信息

Galetzka Danuta, Weis Eva, Kohlschmidt Nicolai, Bitz Oliver, Stein Raimund, Haaf Thomas

机构信息

Institute for Human Genetics, Johannes Gutenberg University, Mainz, Germany.

出版信息

J Cell Biochem. 2007 Apr 1;100(5):1232-9. doi: 10.1002/jcb.21113.

Abstract

Meiosis is the key process for recombination and reduction of the diploid chromosome set to a haploid one. Many genes that have been found in yeast or mouse models to play a role in meiosis are also important for the repair of DNA damage in somatic cells. To study the DNA repair gene transcriptome during male germ cell development, we have developed a specialized cDNA microarray with 181 human genes which are involved in different somatic DNA repair pathways and/or cell cycle control and 45 control house-keeping genes. This DNA repair gene chip was used to quantify the mRNA expression levels in three human testes samples versus a fibroblast RNA pool. Two hundred twenty genes on the chip (including house-keeping genes) showed detectable expression levels in adult testes. Sixty-four DNA repair- and cell cycle-associated genes showed higher expression levels in testicular cells than in mitotically dividing fibroblasts and, therefore, are likely to be implicated in meiosis. The microarray results of 17 genes with increased expression levels were validated with reverse Northern blots or real-time quantitative RT PCR. Systematic analyses of the meiotic DNA repair gene transcriptome may provide new insights into the genetics of male (in)fertility.

摘要

减数分裂是二倍体染色体组重组并减半为单倍体染色体组的关键过程。在酵母或小鼠模型中发现的许多在减数分裂中起作用的基因,对体细胞中DNA损伤的修复也很重要。为了研究男性生殖细胞发育过程中的DNA修复基因转录组,我们开发了一种专门的cDNA微阵列,其中包含181个参与不同体细胞DNA修复途径和/或细胞周期调控的人类基因,以及45个管家基因作为对照。该DNA修复基因芯片用于定量检测三个人类睾丸样本与一个成纤维细胞RNA库中的mRNA表达水平。芯片上的220个基因(包括管家基因)在成人睾丸中显示出可检测的表达水平。64个与DNA修复和细胞周期相关的基因在睾丸细胞中的表达水平高于有丝分裂的成纤维细胞,因此可能与减数分裂有关。通过反向Northern印迹或实时定量RT-PCR验证了17个表达水平升高的基因的微阵列结果。对减数分裂DNA修复基因转录组的系统分析可能为男性生育(不)育的遗传学提供新的见解。

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