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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。
In order to analyze the molecular genetic differences between the anthers of Acanthopagrus schlegelii × Pagrosomus major♂ and the economic traits such as growth and development, which are superior to A.schlegelii, The sequences of the genes PACa M and As Ca M were analyzed in detail using bioinformatics methods in CaM genes of progenies (PA) and black seabream (As). In the meantime, the sequences of PACa M and As Ca M in larvae And second-instar adult fish in different tissues expression characteristics. The results showed that the c DNA of PACa M gene was 1180 bp in length and 1241 bp of As Ca M gene was a 450 bp open reading frame, encoding 149 amino acids with a molecular weight of about 16.84 kD. The isoelectric point was 4.09; Sequence alignment, structural comparison and other analysis showed that PACa M and As Ca M belonged to the Ca M gene family with four EF-hand calcium binding domains. Quantitative analysis showed that CaM was highly expressed in the brains and gonads of larval and adult fish of both species. The expression of PACa M and As Ca M in gill, muscle and gonads of larvae and adult fish was significantly different (P <0.05 ), And there was no significant difference in brain, liver and kidney (P> 0.05). The expression of PACa M was the highest in larval fish and the highest expression of As Ca M was in adult gonads, all of which showed that the Ca M gene in growth and reproduction In the important role. The results provided some basic information for the functional gene expression of the offspring and the parental traits obtained from the cross between the two families.