摘要
本研究通过RACE技术获得了长牡蛎(Crassostrea gigas)Fem-1b和Fem-1c基因cDNA全长序列,并分析了其编码蛋白和系统进化关系。通过RT-PCR技术对采集的长牡蛎周年样品(2015年5月~2016年4月的性腺样品,2015年夏季各组织样品以及不同胚胎幼虫发育时期的样品)进行了表达分析。研究显示,长牡蛎Fem-1b全长2 580bp,编码636个氨基酸;Fem-1c全长2 417bp,编码622个氨基酸。蛋白结构预测显示,两基因分别含有6和7个典型的锚定蛋白重复序列(Anyrin repeat)。RT-PCR结果显示,目的基因在精巢中的表达量显著高于其他组织(P<0.05);Fem-1b在6、7月的精巢中和Fem-1c基因在3、4月的精巢中的表达量显著高于在其他组织中的表达量(P<0.05)。胚胎幼虫表达分析显示,目的基因在胚胎发育早期的表达量显著高于胚胎发育中后期,并在囊胚期达到最高值。我们推测Fem-1b和Fem-1c可能参与了性别决定和性别分化的过程。
We isolated the complete cDNA sequences of Fem-1b and Fem-1c gene of Pacific oyster(Crassostrea giga s)using RACE technique,and analyzed their deduced protein sequences and the phylogenetic relationship of these proteins.In addition,using RT-PCR technique,we did expression analysis in the gonads collected monthly from May 2015 through April 2016,different tissues collected in the summer of 2015 and embryonic larvae at different developmental stages.Results showed that the full length cDNA of Cg-Fem-1b was 2 580 bp encoding 636 amino acids while that of Cg-Fem-1c was 2,417 bp encoding 622 amino acids.The protein structure prediction showed that two genes contained 6 and 7 ankytin repeat motifs,respectively.RT-PCR results showed that the expression of two genes in testis was significantly higher than their expression in other tissues(P<0.05).The expression of Cg-Fem-1b in testis was significantly higher than its expression in other groups in June and July while the expression of Cg-Fem-1c in testis was significantly higher than its expression in other groups in March and April(P<0.05).Two genes highly expressed at the early stages of embryo development in comparison with their expression at middle and late stages,which reached the highest at blastocyst stage(P<0.05).We inferred that Fem-1b and Fem-1c genes may be involved in sex determination and differentiation.
作者
周祖阳
李琪
于红
孔令锋
ZHOU Zu-Yang;LI Qi;YU Hong;KONG Ling-Feng(Thy Key Laboratory of Mariculture(Ocean University of China),Ministry of Education,Qingdao 266003,China)
出处
《中国海洋大学学报（自然科学版）》
CAS
CSCD
北大核心
2018年第6期45-54,共10页
Periodical of Ocean University of China
基金
国家自然科学基金项目(31772843)
泰山学者种业计划专家项目
山东省科技发展计划项目(2016ZDJS06A06)资助~~
