Zhou W, Xu J, Goldberg E
Department of Biochemistry, Northwestern University, Evanston, Illinois 60208.
Biol Reprod. 1994 Sep;51(3):425-32. doi: 10.1095/biolreprod51.3.425.
A clone containing the 5' flanking region of the testis-specific murine lactate dehydrogenase C (Ldhc) gene was isolated from a mouse genomic library. Promoter activity was demonstrated within a 720-bp fragment in testis nuclear extract (TN). Interestingly, the addition of liver nuclear extract (LN) significantly repressed Ldhc promoter activity in the transcription assay system. Sequence analysis of this promoter region revealed several ubiquitous cis-regulatory elements, including one TATA box, one GC box, and two putative CCAAT elements. Analysis of a series of deletion mutants revealed that a 60-bp core promoter sequence was sufficient to direct basal, testis-specific transcription in an in vitro transcription system. A 103-kDa protein in TN and a 65-kDa protein in LN bind to the same palindromic sequences within the 60-bp core promoter region.
从小鼠基因组文库中分离出一个包含睾丸特异性小鼠乳酸脱氢酶C(Ldhc)基因5'侧翼区的克隆。在睾丸核提取物(TN)中的一个720 bp片段内证实了启动子活性。有趣的是,在转录分析系统中添加肝核提取物(LN)可显著抑制Ldhc启动子活性。对该启动子区域的序列分析揭示了几个普遍存在的顺式调控元件,包括一个TATA盒、一个GC盒和两个假定的CCAAT元件。对一系列缺失突变体的分析表明,一个60 bp的核心启动子序列足以在体外转录系统中指导基础的、睾丸特异性的转录。TN中的一种103 kDa蛋白和LN中的一种65 kDa蛋白与60 bp核心启动子区域内的相同回文序列结合。