Venoms, Biological Resources, Molecular Biology

摘要 核心A的目标是维持来自不同芋螺物种的毒液和组织的大量收集以及纯化的毒液肽、合成肽及其类似物的文库。生物材料将主要从菲律宾各地收集。虽然毒液将被用作 纯化的肽，组织将用作mRNA和基因组DNA的来源，用于毒素的分子生物学鉴定。肽将通过生物化学纯化和肽编码核酸序列的分子生物学鉴定来鉴定。从进化论的知识 为了了解芋螺种间的关系，我们将系统地分析芋螺毒液中的特定配体。 利用特异性引物对基因组DNA进行PCR扩增，获得用于锥虫分支分析的遗传标记。 通过从由毒管mRNA构建的cDNA文库中鉴定相应的cDNA来确定完整的毒素（前原成熟毒素）序列。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的毒素库。 分子生物学方法提供了一种从非常有限的生物组织中鉴定潜在配体（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鉴定相应毒素。 核心将继续向科学界提供合成肽和毒液样本。在上一个供资期间提供了100多个实验室。

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.