栉孔扇贝Cf-dmrt4-like基因的克隆、序列特征及表达分析  

Cloning,characterization and expression analysis of Cf-dmrt4-like gene in Chlamys farreri

作　　者：冯政夫[1,2] 邵明瑜[1] 孙大鹏 张志峰[1] 

机构地区：[1]中国海洋大学海洋生物遗传育种教育部重点实验室,山东青岛266003 [2]青岛农业大学,山东青岛266103 [3]辽宁省水产研究所辽宁省海洋水产分子生物学重点实验室,辽宁大连116023

出　　处：《中国水产科学》2010年第5期930-940,共11页Journal of Fishery Sciences of China

基　　金：国家自然科学青年基金面上项目(30901097);山东省自然科学重点基金(Z2008D02)

摘　　要：DMRT家族是一类转录因子,在性别决定与分化、器官形成等早期胚胎发育中起重要的作用。本研究采用简并PCR扩增和cDNA末端快速扩增技术(RACE),从栉孔扇贝(Chlamys farreri)精巢中克隆得到1个全长为2312bp的dmrtcDNA序列,其开放阅读框(Open reading frame,ORF)1110bp,编码369个氨基酸,具有dmrt基因家族共有的DM保守结构域。同源比对和系统进化分析结果显示,其为Cf-dmrt4-like基因。半定量RT-PCR结果显示,该基因从受精卵至匍匐幼虫各发育时期均有表达,卵裂期表达量较高;在雄性成体的鳃和精巢以及雌性成体的外套膜、鳃、肾和闭壳肌中表达,但卵巢中未见表达。qRT-PCR检测不同发育时期的精卵巢,以成熟期精巢表达量最高。由此推测,栉孔扇贝Cf-dmrt4-like基因参与个体的早期发育,并在两性成体中发挥着不同的作用。The dmrt genes encode a large family of transcription factors involved in sex determination,gonad differentiation,and organ formation during early development.The genes were first found in dsx of Drosophila melanogaster and mab-3 of Caenorhabditiselegans,and contain DM domain,a conservative DNA binding motif.In the present study,a complete cDNA of dmrt gene was cloned from testis of Chlamysfarreri by using degenerate PCR and Rapid Amplification of cDNA End（RACE）methods.The full-length of dmrt cDNA is 2312bp,containing a 3＇ untranslated region（UTR）of 1023bp,a 5＇UTR of 179bp,and an open reading fram（eORF）of 1 110 bp which encodes a peptide of 369 amino acids.The putative amino acid sequence shares the DM domain which is common to DMRT family.Phylogenetic tree revealed that the DMRT was clustered with DMRT4,and was most closely related to DMRT4.Therefore,it was named Cf-dmrt4-like gene.Cf-dmrt4-like mRNA was detected by RT-PCR in the developmental stages from fertilized eggs to creeping larva,during which high level expression occured at cleavage embryo stage.The Cf-dmrt4-like gene was also expressed in gill and testis of adult male,as well as in mantle,gill,kidney and adductor muscle of adult female,but not in ovary.The result of qRT-PCR during developmental cycles of both sexual gonads showed high expression of Cf-dmrt4-like gene in testis of mature stage.It could be concluded that Cf-dmrt4-like gene might be involved in early development of C.farreri,and takes on different functions in tissues and gonad development of both male and female.

关 键 词：栉孔扇贝 DMRT基因 全长CDNA 序列特征 组织表达 

分 类 号：S917[农业科学—水产科学]