Venoms, Biological Resources, Molecular Biology

毒液、生物资源、分子生物学

• 批准号: 8380811
• 负责人: PRADIP K BANDYOPADHYAY
• 金额: $ 22.03万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8380811
• 关键词: Automation; Base Sequence; Biochemical; Biocompatible Materials; Biological; Collection; Communities; Complementary DNA; Conotoxin; Conus genus; DNA; Duct (organ) structure; Enzymatic Biochemistry; Family; Freezing; Funding; Genetic Markers; Genomics; High Pressure Liquid Chromatography; Institutes; Knowledge; Laboratories; Libraries; Life; Ligands; Location; Maintenance; Marines; Messenger RNA; Methods; Molecular; Molecular Analysis; Molecular Biology; Nucleic acid sequencing; Peptides; Philippines; Preparation; Procedures; Process; Resources; Sampling; Scanning; Science; Snails; Source; Standardization; System; Tissue Sample; Tissues; Toxin; Universities; Venoms; abstracting; analog; cDNA Library; interest; novel; receptor; small molecule; synthetic peptide

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.

抽象的 核心A的目的是维持来自不同圆锥物种的大量毒液和组织，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的各个位置收集。而毒液将被用作直接来源 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。肽将通过生化纯化和通过分子生物学鉴定肽编码核酸序列来鉴定。从了解进化的知识 圆锥物种之间的关系我们将系统地分析特定配体的毒液。 使用特定引物通过PCR扩增基因组DNA，将获得用于分子分析的临床分析的遗传标记。 完整的毒素（PRO前成熟毒素）序列将通过鉴定从毒液 - 导管mRNA构建的cDNA文库中确定相应的cDNA。具有家族特异性引物的cDNA文库的PCR扩增也将用于确定表达毒素的曲目。 分子生物学方法提供了一种来自非常有限的生物组织的潜在配体（肽）的方法。通过PCR扩增，将使用不同综合毒素超家族的保守引物来鉴定基因组DNA的毒素。 核心将继续向科学界提供合成的肽和毒液样品。在上一个资金期间，提供了100多个实验室。