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Ectopic activation of the transcription promoter for the testis‐specific mouse Pgk‐2 gene on elimination of a cis ‐acting upstream DNA region
Author(s) -
Ando Hiroshi,
Haruna Yoshiya,
Suzuki Misao,
Yamada Shuichi,
Okabe Masaru,
Nakanishi Yoshinobu
Publication year - 2000
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1046/j.1440-169x.2000.00524.x
Subject(s) - biology , microbiology and biotechnology , transcription (linguistics) , promoter , gene , gene expression , northern blot , messenger rna , transgene , southern blot , regulatory sequence , coding region , regulation of gene expression , genetics , philosophy , linguistics
Transgenic mice carrying the coding sequence of β‐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 β‐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 1.4 kb 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, β‐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 β‐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.