半滑舌鳎两种Aquaporin1同源基因的克隆与表达分析

Cloning,Expression and Analysis of Two Aquaporin1 Paralogous Genes in Cynoglossus semilaevis

克隆了半滑舌鳎(Cynoglossus semilaevis)AQP1o和AQP1基因的全长cDNA序列,并对其序列和表达模式进行了分析.结果表明,AQP1ocDNA全长为993 bp,包括120 bp的5′非翻译区,789 bp的开放阅读框,84bp的3′非翻译区,编码262个氨基酸的蛋白质,分子质量为28.3×103.AQP1的cDNA全长为1 335 bp,包括63bp的5′非翻译区,786 bp的开发阅读框,486 bp的3′非翻译区,编码261个氨基酸的蛋白质,分子质量为27.4×103.聚类分析表明半滑舌鳎AQP1o蛋白与金头鲷和塞内加尔鳎等海洋鱼类在卵巢中特异表达的AQP1o聚为一类,而AQP1与非卵巢特异表达的AQP1聚为一类,表明该两类基因为不同类型的AQP1同源基因.RT-PCR分析表明两个基因具有不同的表达模式,AQP1o主要在卵巢中表达,提示其在卵巢中具有特定的生理功能、而AQP1则在雌雄鱼的多种组织中表达.

We cloned the full length cDNA of two AQP1 from Cynoglossus semilaevis and sequenced it. Sequence analysis revealed that the fragment of AQPlo cDNA contained 789 bp open reading frame （ORF）, 120 bp 5＇ untranslated region （UTR） and 84 bp 3＇UTR and that of AQP1 cDNA contained 786 bp open reading frame （ORF）, 63 bp 5＇ untranslated region （UTR） and 486 bp 3＇UTR. The two sequences each encoded 262 amino acids and 261 amino acids, respectively. A phylogenetic tree was constructed with the sequence of AQP1o and AQP1 protein from different species. The AQP1o of Cynoglossus semilaevis was grouped together with some other marine fish＇s AQPlo mainly expressed in ovary. However the AQP1 of Cynoglossus semilaevis was grouped together with the AQP1 not expressed in ovary. This result showed that the two genes were different AQP1 homologous genes. RT-PCR analysis showed that the AQPlo was highly expressed in the ovary and slightly in the kidney of females, but not detected in the other tissues of females and any tissues of males. RT-PCR showed that AQP1 was expressed highly in some different tissues of females and males. This result indicated that the AQPlo might play an important role in ovary.