文献类型：期刊文献

英文题名：Characterization of cDNAs for calmodulin and calmodulin-like protein in the freshwater mussel Hyriopsis cumingii: Differential expression in response to environmental Ca2+ and calcium binding of recombinant proteins

作者：Ren, Gang[1,2];Hu, Xiaolong[1];Tang, Jinyu[1];Wang, Yan[1]

机构：[1]Zhejiang Univ, Coll Anim Sci, Hangzhou 310058, Zhejiang, Peoples R China;[2]Shaoxing Univ, Coll Life Sci, Shaoxing 312000, Peoples R China

年份：2013

卷号：165

期号：3

起止页码：165

外文期刊名：COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY

收录：SCI-EXPANDED(收录号：WOS:000320894200002)、、WOS

基金：We thank Prof. Jinrong Peng and Dr. Yi-hong Guan for assistance in RT-PCR technology. This work was funded by the National Basic Research Program of China (2009CB118705).

语种：英文

外文关键词：Hyriopsis cumingii; HcCaM and HcCaLP; Gene expression; Calcium metabolism; Calcium stimulation

外文摘要：Calmodulin and calmodulin-like protein are two crucial calcium regulators in bivalves. However, molecular characteristics and expression patterns of these genes in the freshwater mussel are poorly understood. In this study, two cDNAs encoding novel calmodulin and calmodulin-like protein (HcCaM and HcCaLP) were cloned and characterized from the freshwater pearl mussel Hyriopsis cumingii. The full-length cDNA of HcCaM was 726 bp, including a 118-bp 5'-untranslated region (UTR), a 447-bp open reading frame (ORF), and a 161-bp 3'-UTR. The 1217-bp HcCaLP cDNA comprised of a 51-bp 5'-UTR, a 447-bp ORF, and a 716-bp 3'-UTR. The potential phosphorylation sites of, Arg(80) and Phe(100) in deduced HcCaM were mutated to Thr(80) and Tyr(100) in HcCaLP. Tissue-specific expression analysis revealed that HcCaM mRNA was prominently expressed in the gill, mantle center, and foot. In contrast, HcCaLP mRNA was mainly expressed in the mantle edge. The recombinant HcCaM and HcCaLP proteins expressed in Escherichia coli showed the typical Ca2+ dependent electrophoretic shift characterization as CaM and differed in the calcium binding affinity. The calcium stimulation test that lasted 5 weeks implied that HcCaM and HcCaLP had differential expression patterns in response to various environmental Ca2+ concentrations (025-1.25 mM). The expression of HcCaM mRNA was up-regulated by low Ca2+ concentration (0.25 mM), and the highest expression of HcCaLP mRNA occurred under Ca2+ concentration of 1 mM. (c) 2013 Elsevier Inc. All rights reserved.