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小鼠睾丸中乳酸脱氢酶C启动子活性的转基因分析。

A transgenic analysis of mouse lactate dehydrogenase C promoter activity in the testis.

作者信息

Kroft Tim L, Li Siming, Doglio Lynn, Goldberg Erwin

机构信息

Department of Biochemistry, Northwestern University, Chicago, Illinois, USA.

出版信息

J Androl. 2003 Nov-Dec;24(6):843-52. doi: 10.1002/j.1939-4640.2003.tb03135.x.

Abstract

Transcription of the mouse testis-specific lactate dehydrogenase c (mldhc) gene is limited to cells of the germinal epithelium. Cloning and analysis of the mldhc promoter revealed that a 100-bp core promoter was able to regulate testis-specific transcription in vitro and in transgenic mice. Surprisingly, expression of the reporter in transgenic testes was limited to pachytene spermatocytes, whereas native LDH-C(4) was detected in pachytene and all later germ cells. To locate additional regulatory sequence that could recapitulate the native LDH-C(4) distribution pattern, we investigated the contribution of 5' and 3' flanking sequences to the regulation of LDH-C(4) expression. We found that transcription factor YY1 binds to the mldhc promoter, that the mldhc 3' untranslated sequence does not permit a postmeiotic expression of a beta-galactosidase reporter in transgenic mice, and that native mldhc mRNA is predominately meiotic, with only a low level of postmeiotic distribution. Our results suggest that the high level of LDH-C(4) in postmeiotic cells results from mRNA and protein stability.

摘要

小鼠睾丸特异性乳酸脱氢酶c(mldhc)基因的转录仅限于生精上皮细胞。对mldhc启动子的克隆和分析表明,一个100bp的核心启动子能够在体外和转基因小鼠中调控睾丸特异性转录。令人惊讶的是,转基因睾丸中报告基因的表达仅限于粗线期精母细胞,而天然的LDH-C(4)在粗线期及所有后期生殖细胞中都能检测到。为了定位能够重现天然LDH-C(4)分布模式的其他调控序列,我们研究了5'和3'侧翼序列对LDH-C(4)表达调控的作用。我们发现转录因子YY1与mldhc启动子结合,mldhc 3'非翻译序列不允许转基因小鼠中β-半乳糖苷酶报告基因在减数分裂后表达,并且天然mldhc mRNA主要在减数分裂期存在,减数分裂后分布水平较低。我们的结果表明,减数分裂后细胞中高水平的LDH-C(4)是由mRNA和蛋白质稳定性导致的。

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