IMPROVED APPROACH FOR IDENTIFYING EXPRESSED SEQUENCES

识别表达序列的改进方法

• 批准号: 3443915
• 负责人: SHERMAN Morton WEISSMAN
• 金额: $ 16.33万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-04-01 至 1995-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3443915
• 关键词: artificial chromosomes; complementary DNA; fusion gene; gene expression; genetic library; genetic promoter element; genetic techniques; high performance liquid chromatography; human genetic material tag; human tissue; messenger RNA; molecular cloning; nucleic acid hybridization; nucleic acid sequence; polymerase chain reaction

We propose to develop methods for preparing libraries of cDNA fragments derived from sequences adjacent to the cap site of mRNAs. These methods will be applied to generate 5' end cDNA libraries from a number of human sources. Methods will be developed to select directly from these libraries those cDNAs encoded by large DNA fragments such as YACs without the need to purify the YAC DNA from the host yeast sequences. These methods would allow selection of cDNAs from arrays of YACs. In addition, the usefulness of the 5' end libraries for obtaining promoter sequences and preparing full-length cDNAs will be evaluated. These libraries will provide an improved approach for detecting expressed sequences and positional cloning of disease genes.

我们建议开发制备cDNA片段文库的方法 来源于与mRNA的帽位点相邻的序列。 这些方法 将应用于从许多人中产生5'端cDNA文库， 源 将开发方法来直接从这些 这些cDNA由大的DNA片段编码，如YAC， 需要从宿主酵母序列中纯化YAC DNA。 这些 方法将允许从YAC阵列中选择cDNA。 此外，本发明还提供了一种方法， 5 ′末端文库用于获得启动子序列的有用性 并对制备全长cDNA进行评价。 这些文库将为检测表达的 序列和疾病基因的定位克隆。