Venoms, Biological Resources, Molecular Biology

摘要 核心A的目标是保持来自不同圆锥体物种的大量毒液和组织的收集 和一个纯化的毒肽、合成肽及其类似物的文库。生物材料将成为 主要从菲律宾的不同地点收集。而毒液将被用作直接来源 纯化的多肽、组织将作为mRNA源和基因组DNA用于分子生物学 毒素的鉴定。多肽将通过生化纯化和分子鉴定两种方法进行鉴定。 编码多肽的核酸序列的生物鉴定。从关于进化的知识 圆锥虫物种之间的关系我们将对特定配体的毒液进行系统分析。 通过对基因组进行聚合酶链式反应扩增，将获得用于锥虫分支分析的遗传标记 使用特定的引物进行DNA扩增。 完整的毒素(早熟毒素)序列将通过相应的鉴定确定 从毒液管道mRNA构建的cDNAs文库中获得cDNA。DNA文库的聚合酶链式反应扩增 还将使用家族特有的引物来确定表达的毒素的谱系。 分子生物学方法提供了一种从非常有限的物质中识别潜在配体(肽)的方法。 大量的生物组织。不同芋螺毒素超家族的保守引物将用于鉴定 通过聚合酶链式反应从基因组DNA中分离出各自的毒素。 CORE将继续向科学界提供合成肽和毒液样本。完毕 在前一个供资期间提供了100个实验室。

Abstract The objective of Core A is to maintain a large collection of venoms and tissues from different Conus species and a library of purified venom peptides, synthetic peptides and their analogs. Biological materials will be collected mainly from various locations in the Phillipines. While the venom will be used as a direct source of purified peptides, tissues will be used as source of mRNA and genomic DNA for the molecular biological identification of toxins. Peptides will be identified both by biochemical purification and by molecular biological identification of peptide-encoding nucleic acid sequences. From a knowledge of the evolutionary relationship among the Conus species we will systematically analyze the venoms for specific ligands. Genetic markers for the cladististic analysis of Conideans will be obtained by PCR amplification of genomic DNA using specific primers. Complete toxin (pre-pro-mature toxin) sequences will be determined by identification of corresponding cDNAs from cDNA libraries constructed from venom-duct mRNA. PCR amplication of cDNA libraries with family-specific primers will also be used to determine the repertoire of expressed toxins. Molecular biological methods offer a means of identification of potential ligands (peptides) from very limited amounts of biological tissue. Conserved primers of different conotoxin superfamilies will be used to identify the respective toxins from genomic DNA by PCR amplification. The core will continue to provide synthetic peptides and venom samples to the scientific community. Over 100 laboratories were supplied during the previous funding period.