CORE--MOLECULAR GENETICS

核心--分子遗传学

• 批准号: 6651267
• 负责人: HANSJUERG ALDER
• 金额: $ 29.68万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6651267
• 关键词: Bax gene /protein; DNA repair; biomedical facility; cadherins; genetic mapping; genotype; growth factor receptors; insulinlike growth factor; loss of heterozygosity; molecular biology information system; molecular genetics; neoplasm /cancer genetics; nucleic acid sequence; oligonucleotides; polymerase chain reaction; transcription factor; transforming growth factors

The role of the Molecular Genetics Core in this project is to provide technical assistance to projects #2 and #3, as well as generate a molecular genetics database to be shared with all the components of this program project. The assistance will involve candidate gene direct sequencing, mouse mapping and genotyping and candidate gene LOH analysis of systemic regions of human chromosomes for Project 2 of the for Project 2. Assistance for Project 3 will involve the analysis of microsatellite instability and sequencing of mismatch repair genes hMSH2 and hMLH1 [when necessary hMSH3, hMSH6 and hPMS2], sequencing of mismatch repair genes hMSH2 and hMSH2 and hMLH1[ when necessary hMSH3, hMSH6 and hPMS2], and sequencing of exons 5-9 of p53 gene and sequencing of codons 12/13 and 61 in the K-ras gene in rectal carcinoma samples. In addition, simple repeat sequences in the coding regions of the Transforming Growth Factor-b1 Type II Receptor (TGF-b1RII), the Insulin-Like Growth Factor II Receptor (IGFII2), BAX, b-catenin and Tcf-4 will be examined to analyze MMR-Pathway and Aneuploid- Pathway alterations present in the tumor samples. The Core contains state of the art molecular biology equipment, including two Applied Biosystems 3948 nucleic acid synthesis and purification systems (48 column capacity each), six Applied Biosystems Prism 377 DNA sequencers. Primary data, analyzed with the assistance of our Shared Computer Facility, will be provided to Principal Investigators of the other projects in clearly understandable formats. Because different steps are involved in the generation of primary data, results will be verified together with other Principal Investigators to eliminate any source of potential errors. By providing primary data concerning the indicated genetic analysis, the Molecular Genetics Core will allow to correlate genetic and clinico-pathological data, thereby giving the opportunity to examine the role of these genes in rectal tumorigenesis. Interaction of the Molecular Genetics Core with all the components involved in this project is a necessary element for the fulfillment of the primary objectives set for the project in its entirety.

分子遗传学核心在这个项目中的作用是为项目#2和#3提供技术援助，并生成一个分子遗传学数据库，与该计划项目的所有组成部分共享。为项目2提供的协助将包括人类染色体系统区域的候选基因直接测序、小鼠定位和基因分型以及候选基因杂合性缺失分析。项目3将包括微卫星不稳定性分析和错配修复基因hMSH2和hMLH1[必要时为hMSH3、hMSH6和hPMS2]的测序，错配修复基因hMSH2和hMSH2和hMLH1[必要时为hMSH3、hMSH6和hPMS2]的测序，以及直肠癌样本中p53基因第5-9外显子的测序和K-ras基因密码子12/13和61的测序。此外，还将检测转化生长因子-b1型受体(TGF-b1RII)、胰岛素样生长因子II受体(IGFII2)、Bax、b-catenin和Tcf-4编码区的简单重复序列，以分析肿瘤样本中存在的MMR途径和非整倍体途径的变化。核心包含最先进的分子生物学设备，包括两个应用生物系统3948核酸合成和纯化系统(每个48柱容量)，六个应用生物系统Prism 377 DNA测序仪。在我们的共享计算机设施的协助下分析的主要数据将以清晰易懂的格式提供给其他项目的首席调查员。由于主要数据的生成涉及不同的步骤，因此将与其他首席调查人员一起核实结果，以消除任何潜在的错误来源。通过提供与指定的遗传分析有关的基本数据，分子遗传学核心将使遗传和临床病理数据相互关联，从而有机会检查这些基因在直肠肿瘤发生中的作用。分子遗传学核心与该项目涉及的所有组件的相互作用是实现为该项目设定的全部主要目标的必要要素。