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Transient expression analysis of the mouse ornithine decarboxylase antizyme haploid‐specific promoter using in vivo electroporation
Author(s) -
Ike Akiko,
Ohta Hiroshi,
Onishi Masayoshi,
Iguchi Naoko,
Nishimune Yoshitake,
Nozaki Masami
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(04)00065-1
Subject(s) - microbiology and biotechnology , electroporation , ornithine decarboxylase antizyme , ornithine decarboxylase , biology , promoter , gene isoform , gene , transgene , gene expression , untranslated region , response element , messenger rna , biochemistry , enzyme
The testicular isoform of the ornithine decarboxylase antizyme (OAZt) gene is expressed exclusively in the haploid spermatids of mice. The 357‐bp region, which includes a TATA‐less promoter and an untranslated region, is sufficient for OAZt gene expression in the spermatids of transgenic mice. In this study, in vivo transient transfection to living mouse testes was used to define the transcriptional regulatory elements of the OAZt gene promoter. We found that the 10‐bp element that contains an initiator (Inr) plays a central role as the core promoter, in combination with a downstream element, while two cyclic adenosine monophosphate‐responsive element (CRE)‐like sites in the upstream region also contribute to promoter activity. The electrophoretic mobility shift assay showed binding of the testis‐specific factors to these elements. Our results show that the in vivo DNA transfer technique enables detailed analysis of haploid germ cell‐specific gene regulation in mice.