Ando H, Haruna Y, Suzuki M, Yamada S, Okabe M, Nakanishi Y
Graduate School of Natural Science and Technology, Kanazawa University, Ishikawa, Japan.
Dev Growth Differ. 2000 Aug;42(4):385-93. doi: 10.1046/j.1440-169x.2000.00524.x.
Transgenic mice carrying the coding sequence of beta-galactosidase, for which expression was driven by various upstream regions including the transcription promoter of the testis-specific mouse Pgk-2 gene, were generated. Expression of beta-galactosidase mRNA driven by the region between nucleotide positions -1404 and +61, with respect to the transcription initiation site numbered +1, was examined by reverse transcription-mediated polymerase chain reaction, blot hybridization and in situ hybridization, and compared with that of endogenous Pgk-2 mRNA. The results revealed that the 1.4kb DNA region is sufficient for determining the organ-specific, developmental stage-specific and spermatogenic stage-specific transcription of the mouse Pgk-2 gene. When the region between -684 and +61 was used to generate transgenic mice, beta-galactosidase mRNA was detectable not only in the testis, but also in other organs such as brain and lung. However, the timing and cell-type specificity of testicular expression of beta-galactosidase mRNA were retained in these mice. Because the region between -1404 and -685 repressed the Pgk-2 promoter in somatic cell-derived cell lines, it is suggested that the organ specificity of Pgk-2 transcription is achieved at least partly by negative regulation.
