RAPD PCR FINGERPRINTING AND MOLECULAR CYTOGENETICS OF HARD TICKS

硬蜱的 RAPD PCR 指纹图谱和分子细胞遗传学

• 批准号: 6204098
• 负责人: SCOTT J GUNN
• 金额: $ 14.19万
• 依托单位: UNIVERSITY OF TEXAS RIO GRANDE VALLEY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6204098
• 关键词: animal population genetics; chemical fingerprinting; chromosomes; cytogenetics; fluorescent in situ hybridization; gel electrophoresis; genetic polymorphism; karyotype; method development; nucleic acid probes; polymerase chain reaction; reagent /indicator; restriction fragment length polymorphism; species difference; statistics /biometry; ticks

The proposed research will investigate the nature and extent of DNA polymorphism in various tick species as identified through agarose gel electrophoresis of random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). Ticks are major vectors of many pathogen organisms which affect humans and their domestic animals. The proposed research will provide the initial investigations of DNA polymorphism, primarily in the moderately repetitive segments, of the tick genome. These areas of the genomes have not been investigated in ticks and offer a potential weather of information on the basic genetics and how characters for the construction and testing of tick phylogeny and systematics. Previously investigated areas of the genome have employed multi locus enzyme electrophoresis and RFLP or DNA sequence data in the nuclear and mitochondrial ribosomal DNA, which are under a different level of selection than the areas most frequently sampled by RAPD-PCR. The research also proposed to construct molecular cytogenetic probes to be used in fluorescent in situ hybridization (FISH) to localized and characterize specific DNA segments within the tick genome. RAPD-PCR data will be collected on all the available tick species using wild caught laboratory reared, or cell lines as the DNA source. Species to be investigated include most species of North American ticks that are known to be major disease vectors. Investigations of each individual will include DNA extraction, RAPD-PCR, agarose gel electrophoresis, and analysis of each genetic relatedness using phenetic and phylogenetic approaches. Molecular cytogenetic probes constructed will be used to investigate the movement and/or lose of material in the genome of North American ticks. The data will be applied to the poorly understood area of tick genetics and extended into tick biology, physiology, systematics, and biological control systems.

这项拟议中的研究将调查DNA的性质和范围， 通过琼脂糖凝胶鉴定不同蜱种的多态性 随机扩增多态性DNA聚合酶链电泳 反应（RAPD-PCR）。蜱是许多病原生物的主要媒介 影响人类和他们的家畜。拟议的研究将 提供了DNA多态性的初步调查，主要是在 蜱虫基因组的中度重复片段。的这些区域 尚未对蜱虫的基因组进行研究， 关于基本遗传学的信息，以及 蜱类发生学和系统学的构建和测试。先前 基因组的研究领域采用了多位点酶 电泳和RFLP或DNA序列数据， 线粒体核糖体DNA，它们在不同的水平下， 选择比地区最频繁采样RAPD-PCR。研究 还提出了构建分子细胞遗传学探针，用于 荧光原位杂交（FISH）定位和表征 蜱虫基因组中的特定DNA片段。 将使用以下方法收集所有可用蜱种属的RAPD-PCR数据： 野生捕获实验室饲养，或细胞系作为DNA来源。物种 包括大多数种类的北美蜱， 已知是主要的疾病媒介。对每个人的调查将 包括DNA提取、RAPD-PCR、琼脂糖凝胶电泳， 使用表型和系统发育分析每个遗传相关性 接近。构建的分子细胞遗传学探针将用于 研究北半球基因组中物质的移动和/或丢失， 美国蜱这些数据将被应用于知之甚少的领域， 蜱遗传学和扩展到蜱生物学，生理学，系统学， 生物控制系统。