盐藻CDPK基因的克隆与生物信息学分析  

Cloning and Bioinformatic Analysis of CDPK Gene of Dunaliella salina

作　　者：张晓琳[1] 柴晓杰[1] 张婷[1] 余祝君[1] 薛飞[1] 

机构地区：[1]大连海洋大学/辽宁省海洋生物资源恢复与生境修复重点实验室/农业部北方海水增养殖重点实验室,辽宁大连116023

出　　处：《核农学报》2013年第4期418-424,共7页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金(30972240);辽宁省教育厅科学技术研究项目(2008T023)

摘　　要：为了研究CDPK基因的结构和功能,应用RT-PCR和RACE方法从盐藻Dunaliella salina中克隆得到CDPK基因(GenBank登录号为JQ964113),并对其进行生物信息学分析。结果表明:盐藻CDPK基因的cDNA全长3 052bp。5'-UTR长70bp;开放阅读框长1650bp,编码549个氨基酸;3'-UTR长1332bp。蛋白表现为弱酸性,且稳定、亲水,二级结构中以无规则卷曲和α-螺旋为主,三级结构同源建模成功。亚细胞定位于叶绿体和细胞核中,无跨膜结构域,无信号肽。蛋白序列中存在4个丝氨酸磷酸化位点、4个苏氨酸磷酸化位点和3个酪氨酸磷酸化位点,蛋白激酶结构域在53～331位氨基酸处,蛋白激酶ATP结合域在59～82位氨基酸处,丝氨酸/苏氨酸蛋白激酶活性域在173～185位氨基酸处,4个EF-hand钙结合域分别在357～385、393～421、429～457、465～493位氨基酸处。进化分析结果表明,克隆的CDPK基因与团藻、衣藻、盐藻Dunaliella tertiolecta的CDPK亲缘关系最近。为进一步研究CDPK基因的功能及探究盐藻耐盐机制的信号转导途径奠定了分子基础。Dunaliella salina CDPK gene （GenBank Accession Number：JQ964113） was cloned by RT-PCR and RACE technology, and further analyzed with bioinformatics approaches. The result showed that the full length of cDNA for CDPK was 3 052bp, which contained a 70bp 5＇ - UTR, a 1 650bp complete open reading frame encoding 549 amino acids, and a 3＇ - UTR of 1 332bp. The corresponding protein was stable, hydrophilic and mildly acidic, with random coil and （x-helix as main components in secondary structure, and the 3D model was successfully constructed. The protein located in chloroplasts and nuclei without transmembrane region and signal peptide. The sequence of protein included four serine phosphorylation sites, four threonine phosphorylation sites and three tyrosine phosphorylation sites. And there was a protein kinase region sitting at 53rd to 331 rd amino acids, a protein kinases ATP-binding region spaning from 59rd to 82rd amino acids, a Serine/Threonine protein kinases active-site consisting of 173rd to 185rd amino acids, and four EF-hand regions distributed in 357rd to 385rd, 393rd to 421rd, 429rd to 457rd, 465rd to 493rd amino acids. Further phylogenic analysis displayed that the protein had the most closest genetic relationship with those CDPK of Volvox carteri f. nagariensis, Chlamydomonas reinhardtii and DunalieUa tertiolecta. The research laid a foundation to further study the CDPK function and the signal transduction pathway of salt resistance mechanism in Dunaliella salina.

关 键 词：盐藻 钙依赖蛋白激酶(CDPKs) CDNA全长 生物信息学分析 

分 类 号：Q943.2[生物学—植物学]