胶州湾浮游桡足类18S核糖体RNA基因序列变异分析

Variation Analysis of 18S Ribosomal RNA Gene Fragments Amplified from Jiaozhou Bay Plankton Copepod

设计了桡足类特异引物,从胶州湾浮游生物混合DNA中选择性扩增了浮游桡足类18S核糖体RNA基因片段,建立了该基因片段变异类型文库。从文库中随机选择83个克隆,对所含的18S核糖体RNA基因片段进行了V .spI和BshNIRFLP分析,发现5种操作分类单元(OTU) ,并对这些OTU的代表克隆进行了测序。分析发现获得的克隆均属于桡足亚纲。根据各OTU覆盖的克隆数计算的胶州湾浮游桡足类遗传多样性指数为0 .96。特定基因序列分析能提供易整合、易比较的多样性数据,也是建立浮游生物群落结构分子生物学剖分方法的基础和形态分类的补充。

A copepod-specific primer was designed and used to amplify the 18S ribosomal RNA gene fragments of Jiaozhou Bay plankton copepod in combination with the universal primer of eukaryotes. A plasmid library containing all possible variants of copepod 18S ribosomal RNA gene fragments was constructed. The fragments were re-amplified from randomly selected 83 recombinants and digested with Vsp I and Bsh NI, yielding 4 and 3 restriction patterns respectively. Restriction fragment length polymorphism analyses with two enzymes revealed 5 operational taxonomy units with their representatives, at least one each, sequenced. All sequences were assigned to copepod in systematic analysis. The genetic diversity index calculated from the abundances of recombinants covered by different OTUs is 0.96. The gene diversity analysis provides not only data that can be easily integrated and compared, but alsoa basis on which the molecular partitioning method of copepod community could be developed.