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大鼠磷酸甘油酸变位酶m基因1.3千碱基对的上游5'区域在转基因小鼠中赋予睾丸和骨骼肌特异性表达。

A 1.3-kb upstream 5' region of the rat phosphoglycerate mutase m gene confers testis and skeletal muscle-specific expression in transgenic mice.

作者信息

Broceño C, Walsh K, Pons G

机构信息

Departament de Ciències Fisiològiques II, Universitat de Barcelona, Barcelona, Spain.

出版信息

Biochem Biophys Res Commun. 1999 Sep 16;263(1):244-50. doi: 10.1006/bbrc.1999.1227.

Abstract

Spermatogenesis is a complex process that occurs in successive mitotic, meiotic and post-meiotic phases and involves a highly regulated selective gene-expression pattern. However, this process has not been well characterised at the gene expression level due to the absence of germinal cell lines. We previously demonstrated that the rat skeletal muscle-specific gene for the glycolytic enzyme phosphoglycerate mutase is also specifically expressed in meiotic and haploid male germ cells from testis (12). To analyse the promoter elements that regulate the transcription of the phosphoglycerate mutase m gene (pgam-m)during spermatogenesis, we developed transgenic mice for a construct containing 1.3 kb from the pgam-m promoter linked to the Escherichia coli LacZ gene. RNA analysis by retrotranscription and PCR amplification of transgene expression showed transcriptional activity in the testis with a pattern during testis development that was identical to the endogenous gene. The transgene was also active in skeletal muscle but not in the adult heart in all the transgenic lines analysed. Collectively, these studies demonstrate that the 1.3 kb pgam-m promoter contains sufficient sequences to specify temporally regulated testis-specific expression as well as skeletal-muscle expression.

摘要

精子发生是一个复杂的过程,发生在连续的有丝分裂、减数分裂和减数分裂后阶段,涉及高度调控的选择性基因表达模式。然而,由于缺乏生殖细胞系,这一过程在基因表达水平上尚未得到充分表征。我们之前证明,糖酵解酶磷酸甘油酸变位酶的大鼠骨骼肌特异性基因也在睾丸的减数分裂和单倍体雄性生殖细胞中特异性表达(12)。为了分析在精子发生过程中调控磷酸甘油酸变位酶m基因(pgam-m)转录的启动子元件,我们构建了转基因小鼠,该构建体包含来自pgam-m启动子的1.3 kb片段,与大肠杆菌LacZ基因相连。通过逆转录和转基因表达的PCR扩增进行RNA分析,结果显示在睾丸中具有转录活性,其在睾丸发育过程中的模式与内源基因相同。在所分析的所有转基因系中,转基因在骨骼肌中也有活性,但在成年心脏中无活性。总的来说,这些研究表明,1.3 kb的pgam-m启动子包含足够的序列来指定时间调控的睾丸特异性表达以及骨骼肌表达。

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