Ectopic activation of the transcription promoter for the testis-specific mouse Pgk-2 gene on elimination of a cis-acting upstream DNA region.
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.
Ando
H
H
Graduate School of Natural Science and Technology, Kanazawa University, Ishikawa, Japan.
Haruna
Y
Y
Suzuki
M
M
Yamada
S
S
Okabe
M
M
Nakanishi
Y
Y
D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008822:Mice, Transgenic; D010735:Phosphoglycerate Kinase / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013091:Spermatogenesis / Q000235:genetics; D013737:Testis / Q000201:enzymology*; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000378:metabolism
D017931:DNA Primers; D012333:RNA, Messenger; D014157:Transcription Factors; D010735:Phosphoglycerate Kinase; D001616:beta-Galactosidase