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为探究Wnt4基因在厚壳贻贝幼虫发育阶段和组织生长过程中的作用,通过RACE技术克隆了厚壳贻贝Wnt4基因c DNA全长序列,该序列全长3342 bp,开放阅读框为1074 bp,编码357个氨基酸。该序列和人(Homo sapiens)、小鼠(Mus musculus)、海胆(Paracentrotus lividus)、栉孔扇贝(Chlamys farreri)和长牡蛎(Crassostrea gigas)等物种的同源性分别为61%、61%、60%、71%和76%。通过实时荧光定量PCR(q RT-PCR)分析Wnt4基因在厚壳贻贝成体多个组织中(外套膜、闭壳肌、鳃、雌雄性腺、足和消化腺)均有表达,其中在外套膜中表达量最高,推测可能与贝壳形成有关;Wnt4基因在厚壳贻贝幼虫发育阶段高表达主要集中在壳顶期,并推测Wnt4基因可能参与了贝壳形态结构发生转变的过程以及某些器官的形成与发育。本研究为进一步开展双壳贝类Wnt基因家族的功能研究提供了理论依据。
In order to explore the role of Wnt4 gene in the growth and development of mussel larvae, the full-length cDNA sequence of Wnt4 gene was cloned by RACE technique. The full length cDNA of Wnt4 gene was 3342 bp with an open reading frame of 1074 bp , Encoding 357 amino acids. The sequence shares 61%, 61%, 70% identity with Homo sapiens, Mus musculus, Paracentrotus lividus, Chlamys farreri and Crassostrea gigas, respectively. 60%, 71% and 76%. The Wnt4 gene was expressed in multiple tissues of adult mussels (mantle, adductor muscle, gill, gonad, gland, foot and digestive gland) by real-time quantitative PCR (q RT-PCR) The highest expression level of Wnt4 may be related to the formation of the shellfish. The high expression of Wnt4 gene in the shellfish mussel larval stage mainly concentrated in the shell top stage, suggesting that the Wnt4 gene may be involved in the transformation of the shell morphological structure as well as in some organs Formation and development. This study provides a theoretical basis for further study on the function of Wnt gene family in bivalve molluscs.