Venoms, Biological Resources, Molecular Biology

摘要 核心A的目标是保存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文库。生物材料将 主要是从菲律宾各地收集的。虽然毒液将被用作 纯化的肽，组织将用作分子生物学的mRNA和基因组DNA的来源。 鉴别毒素。肽将通过生物化学纯化和分子生物学方法鉴定。 肽编码核酸序列的生物学鉴定。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。 通过PCR扩增基因组DNA，获得用于锥虫分支分析的遗传标记。 使用特异性引物的DNA。 完整的毒素（前原成熟毒素）序列将通过鉴定相应的 cDNA来自从毒管mRNA构建的cDNA文库。PCR扩增cDNA文库 家族特异性引物也将用于确定表达的毒素的库。 分子生物学方法提供了一种从非常有限的细胞中鉴定潜在配体（肽）的手段。 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PCR扩增从基因组DNA中提取相应的毒素。 核心将继续向科学界提供合成肽和毒液样本。超过 在上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.