摘要
为分析黑鲷♀×真鲷♂(Acanthopagrus schlegelii♀×Pagrosomus major♂)反交子代在生长、发育等经济性状优于黑鲷亲本(A.schlegelii)的分子遗传差异,本研究克隆了反交子代(PA)与黑鲷(As)的Ca M基因,运用生物信息学方法对基因PACa M与As Ca M的序列进行了详细分析;同时通过荧光定量分析了PACa M与As Ca M在仔鱼及2龄成鱼不同组织的表达特征。研究结果表明,PACa M基因c DNA全长1180 bp,As Ca M基因c DNA全长1241 bp,均具有一个450 bp的开放阅读框,编码149个氨基酸,分子量约16.84 k D,等电点为4.09;序列比对、结构比较等分析表明,PACa M和As Ca M属Ca M基因家族,具有4个EF-hand钙结合功能域。定量分析表明Ca M在两种鱼的仔鱼及成鱼的脑与性腺中有较高表达;PACa M和As Ca M在仔鱼及成鱼的鳃、肌肉及性腺中的表达存在显著差异(P<0.05),在脑、肝及肾中的表达没有明显差异(P>0.05);PACa M在仔鱼中表达量最高,As Ca M在成鱼性腺中表达最高,均显示了Ca M基因在生长与繁殖中的重要作用。研究结果为鲷科属间杂交获得的子代与亲本性状差异的功能基因表达提供一些基础资料。
The hybrid offspring from black porgy(Acanthopagrus schlegeli♀)and red porgy(Pagrosomus major♂)have been successfully farmed in an attempt to develop new marine aquaculture species with optimal target traits.To analyze the molecular mechanism of different growth traits between A.schlegeli parent fish(As)and hybrids(PA),the full-length cDNA sequences of CaM genes(named PACaM and AsCaM)from each were cloned and identified.The expression profiles of CaM genes in six different tissues of two-year-old and30d larvae of F1generation hybrids and pure A.schlegeli individuals were evaluated and compared using real-time quantitative PCR.The PACaM gene is1180base pair(bp)in length and the AsCaM gene is1241bp,both including an open reading frame(ORF)of450bp and encoding a peptide of149amino acids with an estimated molecular mass of16.84kD and a predicted isoelectric point(pI)of4.09.Sequence alignment,structure comparison,and bioinformatics analyses showed that PACaM and AsCaM are members of the CaM family.Functional domain analysis indicated that the protein has four EF-h and calcium-binding domains,with sixteen Ca2+binding sites.PACaM and AsCaM are non-secretory and non-transmembrane proteins,and likely have their functional location in the cytoplasm.Quantification of expressed CaM mRNA showed comparatively high levels in the brain and gonadal tissues of two-year-old fish and in larval fish.PACaM mRNA expression was significantly different from AsCaM in gill,muscle,and gonadal tissue,as well as in larvae,but not significantly different in the brain,liver,or kidney tissues(P>0.05).The stage-specific enrichment of PACaM transcripts in the hybrid larvae,and the highest enrichment of AsCaM in the gonad,may signify an essential role for CaM in growth and development.This study provides a basis for further research into gene function and differing genetic traits between the parent and hybrid offspring of genus crosses of Sparidae.
作者
陈淑吟
李鹏
张志勇
许津
祝斐
贾超峰
王思婷
任忠宏
CHEN Shuyin;LI Peng;ZHANG Zhiyong;XU Jin;ZHU Fei;JIA Chaofeng;WANG Siting;REN Zhonghong(Institute of Oceanology & Marine Fisheries, Nantong 226007, China;Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Sciences, Nanjing Normal University,Nanjing 210023, China)
出处
《中国水产科学》
CAS
CSCD
北大核心
2017年第6期1193-1202,共10页
Journal of Fishery Sciences of China
基金
江苏省海洋与渔业局水产三新工程重大项目(D2015-17)
南通市科技局应用基础研究项目(MS12015071,MS12015070)
关键词
黑鲷
黑鲷×真鲷杂交
CaM基因
分子克隆
MRNA表达
Acanthopagrus schlegeli
Acanthopagrus schlegeli×Pagrosomus major hybridization
calmodulin
gene clone
mRNA expression
