Scieglińska Dorota, Vydra Natallia, Krawczyk Zdzisław, Widłak Wiesława
Department of Tumour Biology, Centre of Oncology, Maria Skłodowska-Curie Memorial Institute, Wybrzeze Armii Krajowej 15, 44-101 Gliwice, Poland.
Biochem J. 2004 May 1;379(Pt 3):739-47. doi: 10.1042/BJ20031842.
The rat Hst70 gene and its mouse counterpart Hsp70.2 are expressed specifically in pachytene primary spermatocytes and spermatids. Here we demonstrate that a 165 bp fragment of the Hst70 gene promoter, containing the T1 transcription start site region, entire exon 1 and 42 bp 5' region of the intron, is sufficient to drive testis-specific expression of the chloramphenicol acetyltransferase reporter gene in transgenic mice with the same developmentally regulated pattern as the endogenous Hsp70.2 gene. We show further that high-level tissue-specific gene expression requires additional sequences localized upstream of the T2 transcription start site. Electrophoretic mobility-shift assay analysis revealed that only testes of juvenile rats, when Hst70 gene expression is repressed, contain proteins that specifically bind to the Oct (octamer) sequence localized directly downstream of the T1 site.
大鼠Hst70基因及其小鼠对应基因Hsp70.2在粗线期初级精母细胞和精子细胞中特异性表达。在此我们证明，Hst70基因启动子的一个165 bp片段，包含T1转录起始位点区域、整个外显子1和内含子的42 bp 5'区域，足以驱动氯霉素乙酰转移酶报告基因在转基因小鼠中进行睾丸特异性表达，且其表达模式与内源性Hsp70.2基因相同，受发育调控。我们进一步表明，高水平的组织特异性基因表达需要位于T2转录起始位点上游的其他序列。电泳迁移率变动分析显示，只有在Hst70基因表达受抑制的幼年大鼠睾丸中，才含有能特异性结合位于T1位点直接下游的Oct（八聚体）序列的蛋白质。
