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太平洋牡蛎(Crassostrea gigas)肌球蛋白必需轻链基因的分子特征与表达谱

Molecular characterization and expression profiles of myosin essential light chain gene in the Pacific oyster Crassostrea gigas.

作者信息

Yu Hong, Li Huijuan, Li Qi

机构信息

Key Laboratory of Mariculture (Ocean University of China), Ministry of Education, Qingdao 266003, China.

Key Laboratory of Mariculture (Ocean University of China), Ministry of Education, Qingdao 266003, China.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2017 Nov;213:1-7. doi: 10.1016/j.cbpb.2017.07.007. Epub 2017 Jul 21.

Abstract

In molluscs, muscle contraction is triggered by a direct binding of Ca to myosin. The myosin essential light chain (MELC) provides the Ca binding site and is of importance for motor function and regulation. In this study, the complete cDNA sequence of MELC gene of the Pacific oyster, Crassostrea gigas, was obtained, and the expression profiles were performed in different tissues and different embryo-larval development stages. The results showed that the full length of C. gigas MELC (CgMELC) cDNA was 659bp, containing a 5'-untranslated region of 73 bp, a 3'-untranslated region of 112bp. The open reading frame encoded a 157 amino acid peptide. The protein sequence of CgMELC contained a conserved EF-hand calcium binding motif, and showed a high sequence identity (68.4-100%) with other bivalves. Quantitative analysis of CgMELC mRNA in tissues indicated that CgMELC was expressed at the highest level in the striated adductor muscle, followed by the smooth adductor muscle and mantle. During the development of embryos and larvae, quantitative analysis and whole-mount in situ hybridization revealed that CgMELC was expressed starting from the blastula stage and abundantly expressed in trochophore and D-shaped larvae, indicating that CgMELC might also be involved in regulation of larval muscle system development. Our data provided valuable information for further researches on the function of MELC and regulation of muscle contraction in oysters.

摘要

在软体动物中,肌肉收缩是由Ca直接与肌球蛋白结合触发的。肌球蛋白必需轻链(MELC)提供Ca结合位点,对运动功能和调节很重要。在本研究中,获得了太平洋牡蛎(Crassostrea gigas)MELC基因的完整cDNA序列,并在不同组织和不同胚胎-幼虫发育阶段进行了表达谱分析。结果表明,太平洋牡蛎MELC(CgMELC)cDNA全长659bp,包含73bp的5'非翻译区、112bp的3'非翻译区。开放阅读框编码一个157个氨基酸的肽。CgMELC的蛋白质序列包含一个保守的EF-手型钙结合基序,与其他双壳类动物具有较高的序列同一性(68.4-100%)。对组织中CgMELC mRNA的定量分析表明,CgMELC在横纹闭壳肌中表达水平最高,其次是平滑肌闭壳肌和外套膜。在胚胎和幼虫发育过程中,定量分析和整体原位杂交显示,CgMELC从囊胚期开始表达,并在担轮幼虫和D形幼虫中大量表达,表明CgMELC可能也参与幼虫肌肉系统发育的调节。我们的数据为进一步研究牡蛎中MELC的功能和肌肉收缩调节提供了有价值的信息。

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