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链状亚历山大藻抑制消减杂交文库的构建及分析
引用本文:史西志,刘兵,贺静静,王梦前,郭皓,苏秀榕,李太武.链状亚历山大藻抑制消减杂交文库的构建及分析[J].水产科学,2009,28(11).
作者姓名:史西志  刘兵  贺静静  王梦前  郭皓  苏秀榕  李太武
作者单位:1. 应用海洋生物技术教育部重点实验室,宁波大学,浙江,宁波,315211
2. 国家海洋环境监测中心,辽宁,大连116023
基金项目:浙江省重大科技攻关项目,浙江省教育厅重点项目 
摘    要:为了筛选链状亚历山大藻特异表达的发光相关基因,应用抑制消减杂交技术,构建了链状亚历山大藻特异表达的cDNA消减文库,共获得500个克隆,通过基因PCR初步筛选表明插入片段的大小主要集中在250~1000 bp,随机挑选10个阳性克隆进行双向测序和序列比对分析,有2个克隆基因片段与已知基因序列高度同源,分别为荧光素结合蛋白和羧化酶/加氧酶,另有8个克隆基因片段在GenBank中未查找到相应的同源基因,可能为未知新基因序列.这些基因的获得为进一步研究链状亚历山大藻特异表达基因,阐明其发光机理及建立特异甲藻的新型检测方法提供重要依据.

关 键 词:链状亚历山大藻  抑制消减杂交  差异表达

Construction and Analysis of Suppression Subtractive Hybridization Library of Dinoflagellate Alexandrium catenella
SHI Xi-zhi,LIU Bing,HE Jing-jing,WANG Meng-qian,GUO Hao,SU Xiu-rong,LI Tai-wu.Construction and Analysis of Suppression Subtractive Hybridization Library of Dinoflagellate Alexandrium catenella[J].Fisheries Science,2009,28(11).
Authors:SHI Xi-zhi  LIU Bing  HE Jing-jing  WANG Meng-qian  GUO Hao  SU Xiu-rong  LI Tai-wu
Abstract:To isolate the bioluminescence specific expressed genes of dinoflagellate Alexandrium catenella(A. catenella), suppression subtractive hybridization was performed to construct the differential expressing cDNA libraries of Alexandrium catenella. A total of 500 cDNA clones were picked. PCR amplification revealed that the length of inserted fragments was mainly from 250 to 1000 base pairs. Ten randomly selected clones after PCR screening were sequenced and analysed in GenBank with Blast search. Two ESTs shared significant identity with the luciferin binding protein and Ribulose-1,5-bisphosphate carboxylase/oxygenase and the other ESTs represent novel genes of Alexandrium catenella with no significant homology in GenBank. These results had provided the foundation for cloning new specific genes of Alexandrium catenella and further studying and establishing the novel detection method for specific dinoflagellata.
Keywords:Alexandrium catenella  suppression subtractive hybridization  differential expression
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