前沟藻18S rDNA序列克隆和分子鉴定分析  

Cloning of 18S rDNA of an Amphidinium sp. and Molecular Identifying Analysis

作　　者：胡乐琴[1] 唐晨[1] 汪卿[1] 陈霁升[1] 何培民[1] 

机构地区：[1]上海海洋大学水产与生命学院,上海201306

出　　处：《生物技术通报》2011年第12期92-95,共4页Biotechnology Bulletin

基　　金：上海市优秀学科带头人项目(08XD1403700);上海市国际合作项目(08540702600);上海市水生生物重点学科资助项目(S30701)

摘　　要：旨在扩增一株未知微藻18S rDNA,并进行序列分析和物种鉴定分析。提取该微藻总DNA,用PCR技术扩增其18S rDNA,对序列进行测序,在GenBank进行序列比对分析,用Clustal X软件构建进化树;用光学显微镜进行形态学分析。经测序其序列长度为1 736 bp,G+C为47%;同源性分析表明,与GenBank中多个强壮前沟藻种的18S rDNA基因序列同源性达99%;与前沟藻属的其它藻种同源性均达95%以上。而与裸甲藻属和环藻属的藻株同源性为85%左右。经光学显微镜观察,该藻具有典型的强壮前沟藻形态。结合该藻18S rDNA序列分析和粗略的形态学分析,推断该藻可能为前沟藻属的强壮前沟藻。微藻形体微小,结构简单,需借助电子显微镜才可准确地从形态学上鉴定其种属,鉴定工作繁琐费时;而利用分子技术,则可能使鉴定工作变得简单快捷。DNA extraction of red tide-forming causative species Amphidinium sp.was performed.PCR technology was used to amplify 18S rDNA.The result of amplification indicated that the length of PCR productions of Amphidinium sp.was 1 736 bp,G＋C was 47%.Then,homology analysis was performed using BLAST in NCBI,and the result indicated that the fragment of 18S rDNA of Amphidinium sp.we cloned had a high homology（99%）with that of Amphidinium carterae,95% homology with any other Amphidinium sp.in GenBank,85% homology with Gymnodinium and Prorocentrum.There was no distinct homology with human genomic and mouse genomic.The results show that,18S rDNA was an effective method for rapid identification of genus,while not quite sure for the species identification.

关 键 词：赤潮藻 前沟藻 18S RDNA 分子鉴定 同源性分析 

分 类 号：Q943.2[生物学—植物学]