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GPX5是一种不依赖硒的谷胱甘肽过氧化物酶编码单拷贝基因,在小鼠附睾中差异表达。

GPX5, the selenium-independent glutathione peroxidase-encoding single copy gene is differentially expressed in mouse epididymis.

作者信息

Zhang Ting, Chabory Eléonore, Britan Aurore, Grignard Elise, Pitiot Olivier, Saez Fabrice, Cadet Rémi, Henry-Berger Joelle, Vernet Patrick, Drevet Joël R

机构信息

CNRS UMR 6247 GReD, Clermont Université, 24, Avenue des Landais, 63177 Aubiére cedex, France.

出版信息

Reprod Fertil Dev. 2008;20(5):615-25. doi: 10.1071/rd08008.

Abstract

Using various molecular approaches, including reverse transcription-polymerase chain reaction (RT-PCR), rapid amplification of cDNA ends-PCR, sequencing, northern and western blotting, we found that the mouse GPX5 gene gives rise to at least three different transcripts that are not expressed at the same levels in the mouse epididymis. In addition to the major GPX5 transcript, we show that minor GPX5 transcripts exist, arising either from precocious termination of transcription or an alternative splicing event within intron 4 of the 5 exon-encoding GPX5 single copy gene. Furthermore, we demonstrate that variants of the GPX5 protein that are correlated with the shorter GPX5 transcripts can be detected in caput epididymidis protein extracts and that the various GPX5 isoforms are subject to differential post-transcriptional maturation processes in the mouse epididymis that essentially involve the addition of O-glycosyl extensions. Using a sensitive poly-A+ mRNA tissue blot, as well as RT-PCR and northern assays, we further show that in addition to being expressed in the epididymis, the GPX5 gene is also expressed, albeit at lower levels, in other tissues of the male genital tract, including the testis and prostate. Finally, we present evidence suggesting that the GPX5 gene is expressed in a temporally regulated manner during mouse embryonic development.

摘要

通过各种分子方法,包括逆转录-聚合酶链反应(RT-PCR)、cDNA末端快速扩增-PCR、测序、Northern印迹和Western印迹,我们发现小鼠GPX5基因产生至少三种不同的转录本,它们在小鼠附睾中的表达水平不同。除了主要的GPX5转录本外,我们还发现存在次要的GPX5转录本,它们要么源于转录的过早终止,要么源于编码GPX5单拷贝基因第5外显子的内含子4内的选择性剪接事件。此外,我们证明在附睾头蛋白质提取物中可以检测到与较短的GPX5转录本相关的GPX5蛋白变体,并且在小鼠附睾中,各种GPX5同工型经历不同的转录后成熟过程,这主要涉及O-糖基延伸的添加。使用灵敏的聚腺苷酸+ mRNA组织印迹以及RT-PCR和Northern分析,我们进一步表明,除了在附睾中表达外,GPX5基因在雄性生殖道的其他组织,包括睾丸和前列腺中也有表达,尽管表达水平较低。最后,我们提供的证据表明,GPX5基因在小鼠胚胎发育过程中以时间调控的方式表达。

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