Identification of calconectin, a calcium-binding protein specifically expressed by the mantle of Pinctada margaritifera
Identification of calconectin, a calcium-binding protein specifically expressed by the mantle of Pinctada margaritifera
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DOI：
10.1016/j.febslet.2006.03.077
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发表时间：
2006-05-01
期刊：
影响因子：
3.5
通讯作者：
Lopez, E
中科院分区：
文献类型：
--
作者：
Duplat, D;Puisségur, M;Lopez, E
珠母贝贝壳中的珍珠层或珍珠母由95-99%的碳酸钙和1-5%的有机基质组成。在本研究中，我们开发了一种独创的技术，通过结合抑制消减杂交(SSH)来鉴定珍珠层形成细胞(NFC)中差异表达的基因，建立了快速扩增cDNAEnds(RACE)-PCR的cDNA消减文库。到目前为止，已获得72个特异的cDNAs序列。其中包括一种含有两个EF-Hand钙结合域的蛋白质，该蛋白质经RACE-PCR扩增后完全测序。NFC和套膜细胞的半定量RT-PCR证实了该基因的特异性表达和SSH方法的特异性。(C)2006年欧洲生化学会联合会。爱思唯尔出版，版权所有。
Nacre or mother-of-pearl in the shell of Pinctada margaritifera is composed of 95-99% calcium carbonate and 1-5%, organic matrix. In this study, we developed an original technique to characterize the genes differentially expressed in nacre-forming cells (NFC) by combining suppression subtractive hybridization (SSH), to establish a cDNA subtractive library, with rapid amplification of cDNA ends (RACE)-PCR. Seventy-two specific cDNA sequences have been obtained so far. These include a protein containing two EF-hand Ca2+-binding domains which was completely sequenced after amplification by RACE-PCR. Its specific expression as well as the specificity of the SSH method was confirmed by semi-quantitative RT-PCR on NFC and mantle cells. (c) 2006 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.