Tsunemoto Kazunobu, Anzai Masayuki, Matsuoka Toshiki, Tokoro Mikiko, Shin Seung-Wook, Amano Tomoko, Mitani Tasuku, Kato Hiromi, Hosoi Yoshihiko, Saeki Kazuhiro, Iritani Akira, Matsumoto Kazuya
Department of Genetic Engineering, Kinki University, Kinokawa, Wakayama, Japan.
Mol Reprod Dev. 2008 Jul;75(7):1104-8. doi: 10.1002/mrd.20863.
We examined the promoter activities of three mouse maternal genes (H1oo, Npm2, and Zar1) in oocytes and pre-implantation embryos, and examined the promoters for cis-acting elements of 5'-flanking region to obtain the best promoter for inducing oocyte-specific gene expression. For the assay, we injected firefly luciferase gene constructs under the control of the promoters into the oocytes and embryos. Each promoter region showed transcriptional activity in oocytes, but not in fertilized embryos. Deletion analysis showed that a putative E-box region at position -72 of the H1oo promoter and at the -180 of the Npm2 promoter were required for basal transcriptional activity in oocytes. Moreover, a putative NBE motif (NOBOX DNA binding elements) (-1796) was shown to enhance basal transcriptional activity of the Npm2 promoter. Thus, the E-box and/or NBE may be key regulatory regions for the expression of the examined maternal genes (H1oo and Npm2) in growing mouse oocytes.
我们检测了三种小鼠母体基因（H1oo、Npm2和Zar1）在卵母细胞和植入前胚胎中的启动子活性，并检测了5'侧翼区域的顺式作用元件的启动子，以获得诱导卵母细胞特异性基因表达的最佳启动子。为了进行检测，我们将萤火虫荧光素酶基因构建体在启动子的控制下注射到卵母细胞和胚胎中。每个启动子区域在卵母细胞中显示出转录活性，但在受精卵中则没有。缺失分析表明，H1oo启动子-72位和Npm2启动子-180位的一个假定E-box区域是卵母细胞基础转录活性所必需的。此外，一个假定的NBE基序（NOBOX DNA结合元件）（-1796）被证明可增强Npm2启动子的基础转录活性。因此，E-box和/或NBE可能是生长中的小鼠卵母细胞中所检测的母体基因（H1oo和Npm2）表达的关键调控区域。
