ISOLATION OF GENES IDENTIFIED BY DEVELOPMENTAL MUTATIONS

通过发育突变鉴定的基因的分离

• 批准号: 3284200
• 负责人: ALLEN D SHEARN
• 金额: $ 16.03万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-05 至 1990-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3284200
• 关键词: DNA; Drosophilidae; alleles; cytogenetics; developmental genetics; endonuclease; gel electrophoresis; gene expression; gene mutation; genetic library; genetic manipulation; genetic markers; genetic transcription; genetic translation; immature animal; molecular cloning; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; plasmids; temperature sensitive mutant; tissue /cell culture

The development of an organism depends on the expression of individual genes and the integration of the products of those genes into units of function. Considerable progress is being made in understanding regulation of gene expression and the direction of future research in that area is becoming apparent. However, virtually nothing is known about the integration into functional units of the products of genes which are essential for various developmental processes. The overall goal of the research in this laboratory is to use the imaginal discs of Drosophila as a model system to discover principles according to which such gene products are integrated into units of function. The potential significance of discovering such principles is that it could facilitate understanding such abnormal human developmental processes as congenital abnormalities and cancer, it could provide insights into increasing the regeneration potential of damaged organs, and it could stimulate new research approaches to the aging process. Many genes which are essential for imaginal disc development in Drosophila have been identified by mutations which cause imaginal disc abnormalities. The analysis of such mutations has led to significant generalizations about imaginal disc development. However, the analysis of such mutations has not, in general, provided useful clues as to the identity of the normal products of those genes nor to their biochemical function. The long-term objective of the proposed research is to identify the primary products of the genes identified by such mutations in order to begin analyzing their biochemical function in imaginal disc development. The specific aims of this proposal are to use two different approaches to isolate those sequences of DNA which correspond to genes identified by mutations. One approach will utilize DNA-mediated transformation of tissue culture cells derived from temperature-sensitive mutants. The other approach will utilize hybrid dysgenesis to induce insertion mutations. Once such DNA sequences are isolated by either method, they will be used as probes to examine the pattern of transcription of the corresponding genes. Those transcripts which are detected will be tested for their ability to be translated in vitro to identify the polypeptides for which they code. The identification of protein products of such genes will be an important step towards understanding the function of those genes, especially when combined with information already obtained about the phenotype caused by mutations in those genes.

生物的发展取决于个体的表达 基因和这些基因产品的整合到 功能。 在理解法规方面正在取得很大进展 基因表达和该领域未来研究的方向是 变得显而易见。 但是，几乎什么都没知道 集成到基因产品的功能单元中 对于各种发展过程至关重要。 总体目标 该实验室的研究是将果蝇的想象盘用作 根据哪些基因产物发现原理的模型系统 被整合到功能单位中。 潜在的意义 发现这样的原则是它可以促进理解这种原则 人类发育过程异常，作为先天性异常和 癌症，它可以提供有关增加再生的见解 器官受损的潜力，它可以刺激新的研究方法 到衰老过程。 许多对于果蝇中想象的椎间盘发育至关重要的基因 已经通过引起想象盘异常的突变来识别。 对此类突变的分析已导致有关 想象中的光盘发育。 但是，对此类突变的分析具有 通常，没有提供有关正常身份的有用线索 这些基因的产物或生化功能。 长期 拟议研究的目的是确定 通过这种突变鉴定的基因，以开始分析其 想象盘发育中的生化功能。 具体目的 该建议是使用两种不同的方法来隔离这些方法 DNA的序列与突变鉴定的基因相对应。 一 方法将利用DNA介导的组织培养细胞的转化 源自温度敏感突变体。 另一种方法将 利用杂交失调诱导插入突变。 曾经这样的DNA 序列是通过两种方法隔离的，它们将用作探针 检查相应基因的转录模式。 那些 检测到的成绩单将被测试 在体外翻译以识别其编码的多肽。 这 鉴定此类基因的蛋白质产物将是一个重要的步骤 要理解这些基因的功能，尤其是当合并时 已经获得有关突变引起的表型的信息 在这些基因中。