| 摘要: |
| Wnt4 作为Wnt 基因家族的重要成员, 在动物的生长发育过程中起重要调控作用。本文利用RACE 技术克隆了长牡蛎Wnt4 基因cDNA 全长序列, 该序列全长1999bp, 开放阅读框为1068bp, 编码355 个氨基酸, 该蛋白序列与人(Homo sapiens)、沙蚕(Platynereis dumerilii)和栉孔扇贝(Chlamys farreri)Wnt4 蛋白的相似性分别为44%、48%和46%。通过荧光定量RT-PCR 分析长牡蛎Wnt4 基因在成体不同组织和不同发育阶段的表达情况, 发现长牡蛎的Wnt4 基因具有广泛的组织表达特点, 在所检测的多种组织中(外套膜、鳃、唇瓣、消化腺、雄性性腺、雌性性腺)均有表达,推测长牡蛎的Wnt4 以信号分子的形式参与多种组织细胞的生命过程; 长牡蛎个体发育过程中Wnt4 基因的高表达主要集中在胚胎发育的早期(桑葚期最高, 原肠胚期次之), 幼贝期该基因的表达量很低, 说明Wnt4 基因可能在早期发育阶段参与了某些器官的形成。 |
| 关键词: 长牡蛎 Wnt4 基因 克隆 表达分析 |
| DOI：10.11693/hyhz20140200062 |
| 分类号: |
| 基金项目:国家重点基础研究发展计划(973 项目), 2010CB126402号; 国家自然科学基金重点项目, 40730845号; 国家高技术研究发展计划(863 项目), 2012AA10A405号; 贝类产业技术体系项目, CARS-48号; 泰山攀登计划项目。 |
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| MOLECULAR CLONING AND EXPRESSION OF WNT4 GENE IN PACIFIC OYSTER CRASSOSTREA GIGAS |
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YANG Mei1,2, XU Fei1, LIU Jun1,2, LI Li1, ZHANG Guo-Fan1
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1.National & Local Joint Engineering Laboratory of Ecological Mariculture, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;2.University of Chinese Academy of Sciences, Beijing 100049, China
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| Abstract: |
| As an important member of the Wnt gene family, Wnt4 plays a pivotal role in development processes of many animals. Using the RACE technique, we obtained a complete cDNA sequence of Wnt4 gene in Pacific oyster (Crassostrea gigas). The full length of this gene was 1999 bpencoding 355 amino acids. Homologous analysis on the amino acids showed that C. gigas Wnt4 was over 40% similar to the most species of Wnt4, including Homo sapiens, Platynereis dumerilii, and Chlamys farreri. QRT-PCR results showed that C. gigas Wnt4 gene was expressed in multiple tissues (mantle, gills, labial palp, digestive gland, male gonad, and female gonad), implying that C. gigas Wnt4 plays various biological roles. Among different developmental stages in C. gigas, the early embryogenesis showed high Wnt4 mRNA expression level (highest in the morula stage, followed by gastrula stage), and lowest in the juvenile stage. Therefore, C. gigas Wnt4 may be involved in the formation of certain organs in early developmental stages. |
| Key words: Crassostrea gigas Wnt4 gene cloning expression analysis |
