Venoms, Biological Resources, Molecular Biology

摘要 Core A 的目标是维护大量来自不同芋螺物种的毒液和组织以及纯化的毒液肽、合成肽及其类似物的库。生物材料将主要从菲律宾各地收集。虽然毒液将被用作直接来源 纯化的肽、组织将用作毒素分子生物学鉴定的 mRNA 和基因组 DNA 的来源。将通过生化纯化和肽编码核酸序列的分子生物学鉴定来鉴定肽。从进化论的知识来看 为了了解圆锥芋属物种之间的关系，我们将系统地分析其毒液中的特定配体。 用于 Conideans 分支分析的遗传标记将通过使用特定引物对基因组 DNA 进行 PCR 扩增来获得。 完整的毒素（前成熟毒素）序列将通过从由毒液导管mRNA构建的cDNA文库中鉴定相应的cDNA来确定。使用家族特异性引物对 cDNA 文库进行 PCR 扩增也将用于确定表达毒素的所有组成部分。 分子生物学方法提供了一种从非常有限的生物组织中识别潜在配体（肽）的方法。不同芋螺毒素超家族的保守引物将用于通过 PCR 扩增从基因组 DNA 中鉴定相应的毒素。 该核心将继续向科学界提供合成肽和毒液样本。上一资助期间向 100 多个实验室提供了资助。

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.