链状亚历山大藻抑制消减杂交文库的构建及分析  

Construction and Analysis of Suppression Subtractive Hybridization Library of Dinoflagellate Alexandrium catenella

作　　者：史西志[1] 刘兵[1] 贺静静[1] 王梦前[1] 郭皓[2] 苏秀榕[1] 李太武[1] 

机构地区：[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

出　　处：《水产科学》2009年第11期675-677,共3页Fisheries Science

基　　金：浙江省重大科技攻关项目(2006C13089);浙江省教育厅重点项目(20051701)

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250～1000bp，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。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 Alexandriurn 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 Alexandriurn catenella with no significant homol- ogy in GenBank. These results had provided the foundation for cloning new specific genes of Alexandriurn catenella and further studying and establishing the novel detection method for specific dinoflagellata.

关 键 词：链状亚历山大藻 抑制消减杂交 差异表达 

分 类 号：S917[农业科学—水产科学]