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海带CRY-DASH基因的克隆与转录表达分析
李璐
中国科学院海洋研究所
摘要:
蓝光是海水中能传播的主要光谱,其对海带的生长和发育起重要作用。隐花色素/光裂解酶家族(Cryptochrome/Photolyase Family, CPF)是一类广泛存在的能被蓝光诱导激活的蛋白家族。CRY-DASH (Cryptochrome-Drosophila, Arabidopsis, Synechocystis, Human)是该家族的新成员。本研究利用cDNA末端快速克隆(RACE)技术,获得海带CRY-DASH基因(SjCRY-DASH)全长序列,结构分析发现,其ORF区长1779 bp,编码592个氨基酸。进行氨基酸同源序列比对,其与其它藻类和高等植物间存在两个重要辅基MTHF和FAD结合的保守域。通过不同光质照射诱导海带幼孢子体,我们发现蓝光、白光诱导1 h后,均能使SjCRY-DASH转录水平上升,且SjCRY-DASH对蓝光的响应更强烈。本研究结果为研究大型褐藻-海带中CRY-DASH受光诱导调控功能打下基础。
关键词:  蓝光  海带  CRY-DASH  孢子体
DOI:10.11759/hykx20170222003
分类号:X55
基金项目:国家海洋公益性行业科研专项(201405040);青岛海洋科学与技术国家实验室计划项目(2015ASKJ02)
Cloning and transcriptional analysis of CRY-DASH from Saccharina japonica (Laminariales, Phaeophyceae)
Li Lu
Institute of Oceanology, Chinese Academy of Sciences
Abstract:
Blue light is predominant below seawater surface, and it plays a critical role in regulating growth and development of Saccharina japonica. The cryptochrome/photolyase family (CPF) is a diversified protein family which can be induced and activated by blue light. CRY-DASH (Cryptochrome-Drosophila, Arabidopsis, Synechocystis, Human) is a new member of this family. Here we characterize the full-length cDNA of CRY-DASH from S. japonica (designated as SjCRY-DASH) using rapid-amplication of cDNA ends (RACE). The open reading frame (ORF) length of SjCRY-DASH is 1779 bp, encoding 592 amino acids. By multiple alignments of protein sequences, conserved residues that interact with two crucial cofactors, MTHF and FAD, are detected between SjCRY-DASH and several DASH-type proteins of other algae and higher plant. Juvenile sporophytes are induced by light of different wavelengths. Both blue light and white light upregulate the transcriptional level of SjCRY-DASH in 1 h, though stronger response to blue light was found for SjCRY-DASH than white light. Our results lay foundation for future study of light induction and regulation on CRY-DASH of S. japonica.
Key words:  blue light  Saccharina japonica  CRY-DASH  sporophytes
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