SGER: DNA Thermal Profiling: A New Technique in Conservation Genetics

SGER：DNA 热分析：保护遗传学的新技术

• 批准号: 9528152
• 负责人: Michael Perlin
• 金额: $ 2.8万
• 依托单位: University of Louisville Research Foundation Inc
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-12-01 至 1996-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9528152&HistoricalAwards=false
• 关键词: SGER; DNA; Thermal; Profiling; New

9528152 PERLIN For purposes of wildlife conservation it is important to be able to identify individual animals to be used in captive breeding programs and to assess populations of endangered species. Several methods have been developed to produce molecular "fingerprints" for such analyses. With the rapid decrease of biodiversity, quick and effective methods of assaying biological molecules (such as DNA) are important. Dr. Perlin has developed a simple technique called DNA thermal profiling. The technique employs analysis of how an organism's DNA melts. Initial evidence indicates that each individual or species may have a unique melting profile. This technique circumvents the more tedious and costly methods currently in use in many labs. The new method is also technically simpler and data can be generated in less time. Furthermore, the amount of DNA required is small (100-200 ng) and total genomic DNA is used as opposed to small, selected portions. Thus, it has the advantage of detecting and utilizing microheterogeneities throughout the test organisms' genomes. It may thus produce a true DNA "fingerprint" of the organism. Dr. Perlin has applied DNA thermal profiling to the study of redwing blackbirds, Cuban crocodiles, and alligators. He proposes to determine the genetic basis of the technique and thereby further strengthen its validity. The data obtained from animals with well-established pedigrees (e.g., racehorses and beagles) will be examined for Mendelian patterns of inheritance. In addition, populations of endangered animals maintained in zoos or breeding facilities will be studied to determine whether this technique can be used effectively for such population analyses. As much as possible, all data obtained with DNA thermal profiling will be compared with that obtained by already-established methods for DNA fingerprinting and population studies. Dr. Perlin is primarily interested in studying the limits of this new and potentially exciting method in environmental biology. If the method is robust, the results of this study could demonstrate that DNA thermal profiling is a quick and simple method for DNA fingerprinting with particular significance for conservation biologists and captive breeding programs.

对于野生动物保护而言，能够识别用于圈养繁殖计划的个体动物和评估濒危物种的种群是很重要的。已经开发了几种方法来产生用于这种分析的分子“指纹”。随着生物多样性的急剧减少，快速有效地分析生物分子（如DNA）的方法变得非常重要。珀林博士开发了一种简单的技术，叫做DNA热分析。这项技术通过分析生物体的DNA是如何融化的。初步证据表明，每个个体或物种可能都有独特的融化剖面。这项技术绕过了目前许多实验室使用的更繁琐和昂贵的方法。新方法在技术上也更简单，可以在更短的时间内生成数据。此外，所需的DNA量很小（100-200 ng），并且使用的是全部基因组DNA，而不是一小部分选定的DNA。因此，它具有检测和利用整个被试生物基因组的微异质性的优势。因此，它可能产生生物体的真正DNA“指纹”。Perlin博士将DNA热分析应用于红翼黑鸟，古巴鳄鱼和短吻鳄的研究。他建议确定该技术的遗传基础，从而进一步加强其有效性。从家谱确定的动物（如赛马和小猎犬）获得的数据将用于孟德尔遗传模式的检验。此外，还将对动物园或繁殖设施中濒危动物的种群进行研究，以确定这项技术是否能有效地用于此类种群分析。通过DNA热分析获得的所有数据将尽可能地与已经建立的DNA指纹和人口研究方法获得的数据进行比较。Perlin博士主要感兴趣的是研究这种可能令人兴奋的新方法在环境生物学中的局限性。如果该方法是可靠的，那么本研究结果将证明DNA热谱分析是一种快速简便的DNA指纹识别方法，对保护生物学家和圈养育种计划具有特殊意义。