Marine Biotech Fellowship: A Molecular Analysis of the Phylum Ctenophora

由于无法区分一个物种的遗传结构，远洋生物地理学受到了限制。这些问题包括当地人口的不断流动、未知的海洋环流模式、海洋的三维方面以及海洋取样固有的困难。该项目将使用现代分子技术来研究栉水母门的成员，以帮助阐明这一未被充分研究的生物群体的分类。栉水母在所有的海洋中都很丰富，并且出现在水柱的所有深度。栉水母的分布通常是不规则的，这些不规则的分布在时间和地点上都有波动。分子生物学家开发的技术（如同工酶和核酸分析）适用于海洋科学，为海洋科学家研究海洋种群分布提供了新的工具。这些技术已用于渔业，但尚未应用于胶状浮游动物，尽管这一群体的重要性和丰富性。有许多栉水母物种出现在蒙特利湾地区，其中大部分仍未被描述。本项目将包括：1。蒙特雷湾浅水和深水标本采集及基因组DNA分离；2. 利用聚合酶链反应扩增核编码的小核糖体亚基基因（ssRNA）、间隔区和线粒体基因组的某些DNA片段；3. 用限制性内切酶分析扩增的DNA，以检测任何存在的限制性内切片段长度多态性。直接从PCR产物或克隆产物中对扩增DNA进行测序；5. 序列分析将使用DOTPLOT、BESTFIT和GAP/RANDOMIZATION；6. 利用分子资料和经典资料（形态学和自然史）的结合澄清栉虫门的分类。ssRNA序列的使用将对更高的分类水平最有用，而来自间隔区或mtDNA区域的序列信息将对研究种内变异有用。

Pelagic biogeography has been limited by the inability to distinguish genetic structure of a species. The problems include constant movement of local populations, unknown oceanic circulation patterns, the 3-dimensional aspect of the ocean and the difficulties inherent with oceanic sampling. This project will examine members of the Phylum Ctenophora using modern molecular techniques to help elucidate the taxonomy of this understudied group of organisms. Ctenophores are abundant in all the oceans and occur at all depths in the water column. Distribution of ctenophores is often patchy and these patches can fluctuate in time and location. The techniques developed by molecular biologists (e.g. allozymes and nucleic acid analyses) are appropriate for the marine sciences and represent novel tools for the marine scientist to examine the distribution of marine populations. These techniques have been utilized for fisheries but have not been applied to gelatinous zooplankton despite the importance and abundance of this group. There are a number of ctenophore species that occur in the Monterey Bay area, the majority of which are still undescribed. This project will consist of:: 1. Collection of specimens and the isolation of genomic DNA focused on Monterey Bay with shallow-water and deep- water collections; 2. Use of the Polymerase Chain Reaction to amplify the nuclear encoded small ribosomal subunit genes (ssRNA), spacer regions, and certain DNA segments of the mitochondrial genome; 3. Analyze the amplified DNA with restriction enzymes in order to detect any Restriction Fragment Length Polymorphisms that are present: 4. Sequencing of the amplified DNA either directly from the PCR product or after cloning the product; 5. Analysis of the sequences will be done using DOTPLOT, BESTFIT, and GAP/RANDOMIZATION; 6. Clarification of the taxonomy of the Phylum Ctenophora using a combination of the molecular data and classical data (morphology and natural history). The use of the ssRNA sequences will be most useful for the higher taxonomic levels while the sequence information from the spacer region or mtDNA regions will be useful for examining the variation within species.