Department of Biology, Providence College, Providence, Rhode Island, USA.
Cytoskeleton (Hoboken). 2011 Oct;68(10):566-77. doi: 10.1002/cm.20531. Epub 2011 Oct 11.
The squid giant axon and synapse are unique systems for studying neuronal function. While a few nucleotide and amino acid sequences have been obtained from squid, large scale genetic and proteomic information is lacking. We have been particularly interested in motors present in axons and their roles in transport processes. Here, to obtain genetic data and to identify motors expressed in squid, we initiated an expressed sequence tag project by single-pass sequencing mRNAs isolated from the stellate ganglia of the Woods Hole Squid, Loligo pealei. A total of 22,689 high quality expressed sequence tag (EST) sequences were obtained and subjected to basic local alignment search tool analysis. Seventy six percent of these sequences matched genes in the National Center for Bioinformatics databases. By CAP3 analysis this library contained 2459 contigs and 7568 singletons. Mining for motors successfully identified six kinesins, six myosins, a single dynein heavy chain, as well as components of the dynactin complex, and motor light chains and accessory proteins. This initiative demonstrates that EST projects represent an effective approach to obtain sequences of interest.
鱿鱼巨轴突和突触是研究神经元功能的独特系统。虽然已经从鱿鱼中获得了一些核苷酸和氨基酸序列，但缺乏大规模的遗传和蛋白质组学信息。我们特别感兴趣的是轴突中存在的马达及其在运输过程中的作用。在这里，为了获得遗传数据并鉴定鱿鱼中表达的马达，我们通过对来自伍兹霍尔鱿鱼（Loligo pealei）星状神经节的 mRNA 进行单轮测序，启动了一个表达序列标签项目。总共获得了 22689 条高质量的表达序列标签 (EST) 序列，并进行了基本局部比对搜索工具分析。这些序列中有 76%与国家生物信息学数据库中的基因相匹配。通过 CAP3 分析，该文库包含 2459 个重叠群和 7568 个单序列。马达的挖掘成功地鉴定了六个驱动蛋白、六个肌球蛋白、一个单独的动力蛋白重链，以及动力蛋白复合物的组件，以及马达轻链和辅助蛋白。这一举措表明 EST 项目是获得感兴趣序列的有效方法。
