摘要
为了研究IGF1R基因在泥蚶生长、发育过程中的调控作用,本研究基于泥蚶转录组文库的EST,利用SMART RACE技术首次克隆获得泥蚶IGF1R基因(Tg-IGF1R)c DNA全长序列,该序列全长为5793 bp,开放阅读框为4638 bp,编码1546个氨基酸。经生物信息学分析发现,该基因5′端非编码区为810 bp,3′端非编码区为345 bp,Tg-IGF1R蛋白分子量为176.01 ku,等电点为6.04,包含4个Ⅲ型纤粘连蛋白结构域,1个半胱氨酸富集区和1个酪氨酸激酶结构域,属于亲水性跨膜蛋白质。氨基酸多重比对显示Tg-IGF1R蛋白序列与人、非洲爪蟾、斑马鱼的相似性分别为45.2%、45.6%和45.4%。利用q RT-PCR技术检测了Tg-IGF1R基因在泥蚶成体不同组织及幼体不同发育时期的表达情况,发现该基因具有广泛的组织表达性,在所检测的6个组织(血液、闭壳肌、斧足、鳃、内脏团和外套膜)中均有不同程度的表达,其中血液中的相对表达量最高,并与其余5个组织间存在显著性差异,推测Tg-IGF1R以信号分子的形式参与多种组织细胞的生命过程。Tg-IGF1R基因在泥蚶个体发育过程中的9个时期均有表达,且高表达主要集中在原肠胚期,其次是担轮幼虫期和稚贝期,说明Tg-IGF1R基因可能在早期发育阶段参与了某些器官的形成。
In order to explore the function of IGF1 R during the process of the growth and development in Tegillarca granosa,Tg-IGF1 R c DNA was cloned by SMART RACE（Rapid Amplification of c DNA Ends）techniques on the basis of c DNA library of T.granosa. The full length c DNA of Tg-IGF1 R was 5793 bp,and its complete ORF（Open Reading Frame） was 4638 bp encoding 1546 amino acids. Bioinformatics analysis showed that the sequence of Tg-IGF1 R contains a 5′-untranslated region（UTR） of 810 bp,and a 3′-UTR of 345 bp. The predicted molecular mass of the deduced amino acid of Tg-IGF1 R was 176.01 ku,and the theoretical isoelectric point was 6.04. Tg-IGF1 R protein consists of four fibronectin type（FN3） domains,one furin like cysteine-rich（FU） domain and Tyrosine kinase（Tyr Kc） domain,belonging to transmembrane and hydrophilic protein.Homologous analysis on the amino acids showed that IGF1 R of T. granosa shared more than 40% similarity with the most species of IGF1 R,including Homo sapiens,Xenopus laevis,and Danio rerio. q RT-PCR（Real-time quantitative PCR） results indicated that Tg-IGF1 R gene was expressed in 6 tissues（blood,adductor muscle,foot,gill,visceral mass and mantle.）,and the expression of blood was significantly higher than other tissues,which implied that Tg-IGF1 R plays various biological roles. The results of relative expression in 9 development stages revealed that the expression of Tg-IGF1 R gene showed the highest expression in the gastrulae stage,and followed by trochophore and juvenile clams stages. Therefore,Tg-IGF1 R may be involved in the formation of certain organs in early developmental stages.
出处
《水产学报》
CAS
CSCD
北大核心
2017年第1期40-51,共12页
Journal of Fisheries of China
基金
国家现代贝类产业技术体系专项(CARS-48)
国家水产种质资源平台项目(2015DKA30470)
浙江省自然科学基金重点项目(LZ12C19001)
浙江省重中之重学科自设课题(ZS2015012)~~
