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A testis-specific gene encoding a nuclear high-mobility-group box protein located in elongating spermatids.

机译:睾丸特异性基因编码位于伸长的精子细胞中的核高迁移率族盒蛋白。

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摘要

A cDNA encoding a DNA-binding protein has been isolated by screening a mouse testicular expression cDNA library with a concatemer of a 12-bp putative protein-binding element present in the promoter of the testis-specific gene PGK-2. Sequence analysis of the isolated cDNA indicated the presence of an open reading frame that encodes a protein with two conserved DNA-binding motifs known as the high-mobility-group (HMG) boxes. Northern (RNA) blot analysis demonstrated that expression of the gene is restricted to the postpuberal testis. The DNA-binding activity and sequence specificity of the recombinant HMG protein were confirmed by DNA mobility shift assay using the initial concatemer of the PGK-2 promoter element as a probe as well as the wild-type or mutated versions of the 12-bp element within its natural sequence context. Immunocytochemical staining of adult testis sections with polyclonal antisera recognizing this recombinant HMG protein demonstrated that it is located predominantly in the nuclei of elongated spermatids at steps 9 and 10. These results suggest that this novel HMG box protein gene may be involved in the regulation of gene expression of the haploid male genome. The gene from which the cDNA was derived has been termed testis-specific HMG (tsHMG).
机译:通过用存在于睾丸特异性基因PGK-2启动子中的12 bp推定蛋白结合元件的串联体筛选小鼠睾丸表达cDNA文库,已分离出编码DNA结合蛋白的cDNA。分离的cDNA的序列分析表明存在一个开放阅读框,该阅读框编码具有两个保守的DNA结合基序的蛋白质,称为高迁移率族(HMG)盒。 Northern(RNA)印迹分析表明该基因的表达仅限于青春期后的睾丸。重组HMG蛋白的DNA结合活性和序列特异性通过PGK-2启动子元件的初始连接体作为探针以及12 bp元件的野生型或突变形式通过DNA迁移转移测定法得以证实在其自然序列范围内。用识别该重组HMG蛋白的多克隆抗血清对成人睾丸切片进行免疫细胞化学染色显示,该蛋白主要位于第9步和第10步细长精子的细胞核中。这些结果表明,该新型HMG box蛋白基因可能参与了该基因的调控单倍体男性基因组的表达cDNA的衍生基因已被称为睾丸特异性HMG(tsHMG)。

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