中国水产科学2017，Vol.24Issue(6)：1193-1202,10.DOI:10.3724/SP.J.1118.2017.17027
黑鲷♀×真鲷♂反交子代与黑鲷的CaM基因克隆与mRNA表达分析
Molecular cloning, mRNA expression, and characterization of calm-odulin genes in hybrids of Acanthopagrus schlegeli (♀)×Pagrosomus major (♂)
摘要
Abstract
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 and 30 d larvae of F1 generation hybrids and pure A. schlegeli individuals were evaluated and compared using real-time quantitative PCR. The PACaM gene is 1180 base pair (bp) in length and the AsCaM gene is 1241 bp, both including an open reading frame (ORF) of 450 bp and encoding a peptide of 149 amino acids with an estimated molecular mass of 16.84 kD and a predicted isoelectric point (pI) of 4.09. Sequence alignment, structure comparison, and bioinfor-matics analyses showed that PACaM and AsCaM are members of the CaM family. Functional domain analysis in-dicated 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 cyto-plasm. 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.关键词
黑鲷/黑鲷×真鲷杂交/CaM基因/分子克隆/mRNA表达Key words
Acanthopagrus schlegeli/Acanthopagrus schlegeli×Pagrosomus major hybridization/calmodulin/gene clone/mRNA expression分类
生物科学引用本文复制引用
陈淑吟,李鹏,张志勇,许津,祝斐,贾超峰,王思婷,任忠宏..黑鲷♀×真鲷♂反交子代与黑鲷的CaM基因克隆与mRNA表达分析[J].中国水产科学,2017,24(6):1193-1202,10.基金项目
江苏省海洋与渔业局水产三新工程重大项目(D2015-17) （D2015-17）
南通市科技局应用基础研究项目(MS12015071, MS12015070). （MS12015071, MS12015070）
