Molecular Cloning and Identification of the Transcriptional Regulatory Domain of the Goat Neurokinin B Gene <i>TAC3</i>
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- SUETOMI Yuta
- Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
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- MATSUDA Fuko
- Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
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- UENOYAMA Yoshihisa
- Laboratory of Reproductive Science, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
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- MAEDA Kei-ichiro
- Laboratory of Animal Breeding, Department of Veterinary Medical Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan
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- TSUKAMURA Hiroko
- Laboratory of Reproductive Science, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
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- OHKURA Satoshi
- Laboratory of Animal Production Science, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
Bibliographic Information
- Other Title
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- Molecular Cloning and Identification of the Transcriptional Regulatory Domain of the Goat Neurokinin B Gene TAC3
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Abstract
Neurokinin B (NKB), encoded by TAC3, is thought to be an important accelerator of pulsatile gonadotropin-releasing hormone release. This study aimed to clarify the transcriptional regulatory mechanism of goat TAC3. First, we determined the full-length mRNA sequence of goat TAC3 from the hypothalamus to be 820 b, including a 381 b coding region, with the putative transcription start site located 143-b upstream of the start codon. The deduced amino acid sequence of NKB, which is produced from preproNKB, was completely conserved among goat, cattle, and human. Next, we cloned 5’-upstream region of goat TAC3 up to 3400 b from the translation initiation site, and this region was highly homologous with cattle TAC3 (89%). We used this goat TAC3 5’-upstream region to perform luciferase assays. We created a luciferase reporter vector containing DNA constructs from –2706, –1837, –834, –335, or –197 to +166 bp (the putative transcription start site was designated as +1) of goat TAC3 and these were transiently transfected into mouse hypothalamus-derived N7 cells and human neuroblastoma-derived SK-N-AS cells. The luciferase activity gradually increased with the deletion of the 5’-upstream region, suggesting that the transcriptional suppressive region is located between –2706 and –336 bp and that the core promoter exists downstream of –197 bp. Estradiol treatment did not lead to significant suppression of luciferase activity of any constructs, suggesting the existence of other factor(s) that regulate goat TAC3 transcription.
Journal
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- Journal of Reproduction and Development
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Journal of Reproduction and Development 59 (5), 463-469, 2013
The Society for Reproduction and Development
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Details
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- CRID
- 1390282681312443520
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- NII Article ID
- 130003361048
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- NII Book ID
- AA10936678
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- COI
- 1:STN:280:DC%2BC3sjmvVOlsQ%3D%3D
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- ISSN
- 13484400
- 09168818
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- NDL BIB ID
- 024943079
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- PubMed
- 23812498
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
- KAKEN
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- Abstract License Flag
- Disallowed