Cloning and expression analysis of IGF1R gene in Tegillarca granosa
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Key Laboratory of Aquatic Germplasm Resource of Zhejiang,Zhejiang Wanli University,Ningbo,Key Laboratory of Aquatic Germplasm Resource of Zhejiang,Zhejiang Wanli University,Ningbo,College of Fisheries and Life Sciences,Shanghai Ocean University,Key Laboratory of Aquatic Germplasm Resource of Zhejiang,Zhejiang Wanli University,Ningbo,Key Laboratory of Aquatic Germplasm Resource of Zhejiang,Zhejiang Wanli University,Ningbo

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    Abstract:

    In order to explore the function of IGF1R during the process of the growth and development in Tegillarca granosa, Tg-IGF1R cDNA was cloned by SMART RACE (Rapid Amplification of cDNA Ends) techniques on the basis of cDNA library of T.granosa. The full length cDNA of Tg-IGF1R 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-IGF1R 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-IGF1R was 176.01 ku, and the theoretical isoelectric point was 6.04. Tg-IGF1R protein consists of four fibronectin type Ⅲ(FN3) domains, one furin like cysteine-rich (FU) domain and Tyrosine kinase (TyrKc) domain, belonging to transmembrane and hydrophilic protein. Homologous analysis on the amino acids showed that IGF1R of T. granosa shared more than 40% similarity with the most species of IGF1R, including Homo sapiens, Xenopus laevis, and Danio rerio. qRT-PCR (Real-time quantitative PCR) results indicated that Tg-IGF1R 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-IGF1R plays various biological roles. The results of relative expression in 9 development stages revealed that the expression of Tg-IGF1R gene showed the highest expression in the gastrulae stage, and followed by trochophore and juvenile clams stages. Therefore, Tg-IGF1R may be involved in the formation of certain organs in early developmental stages.

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REN Fuzhen, YAO Hanhan, QIAN Xuejun, LIN Zhihua, DONG Yinghui. Cloning and expression analysis of IGF1R gene in Tegillarca granosa[J]. Journal of Fisheries of China,2017,41(1):40~51

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History
  • Received:December 13,2015
  • Revised:May 26,2016
  • Adopted:November 15,2016
  • Online: January 19,2017
  • Published:
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