CORE--CYTOGENETICS

核心--细胞遗传学

• 批准号: 5209113
• 负责人: JOHN R MCGILL
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5209113
• 关键词: biomedical facility; cytogenetics; dissection; fluorescence microscopy; fluorescent in situ hybridization; genetic mapping; hybrid cells; karyotype; neoplasm /cancer genetics; neoplastic cell; tissue /cell culture

The purpose of the Cytogenetics Laboratory Shared Resource is to provide cytogenetic analysis, gene mapping and chromosomal microdissection services for members of the San Antonio Cancer Institute. The technical expertise and equipment required for cytogenetic analysis, gene mapping and chromosomal microdissection make it cost prohibitive and undesirable for independent investigators to perform these studies in their own laboratories. The past options for Cancer Center members have been to contract for these services commercially or collaborate with clinical services laboratories which are concerned with patient sample analysis. The objective of this core will be to provide San Antonio Cancer Institute members affordable comprehensive cytogenetic studies for research specimens for the actual costs of the services. a. Cytogenetic analysis of tumor specimens and cell lines Chromosomal analysis of cell lines or tumor specimens: Techniques which will be offered include G-banding (trypsin, Wright's stain, Borate), Q- banding, C- and R-banding, G-11, high resolution banding. 10 - 15 metaphase spreads will be analyzed and a karyotype will be produced. The laboratory is set up for analysis of chromosomes from a variety of tissue types including: tissue cultured cells (adherent or suspension cells) peripheral blood cells, bone marrow and solid tumor. Chromosomal analysis of human-rodent somatic cell hybrid cells will also be performed. b. Chromosomal in situ hybridization Techniques offered include (a) FISH (fluorescent in situ hybridization) analysis for chromosome labeling or gene mapping with cosmids and (b) radiolabeled in situ hybridization for gene mapping with small inserts. c. Chromosome microdissection and microcloning (CMM) Chromosome microdissection for the molecular analysis of specific chromosome regions will be offered. This recently described technique allows small pieces of chromatin to be selectively scrapped from chromosomes. This method provides clones for the starting points for molecular analysis from specific chromosome regions linked to specific diseases (Rohme et al., 1984; Ludecke et al., 1989). There are many specific chromosomal regions of interest associated with neoplasia that this procedure might prove invaluable for the generation of new genetic tumor markers. CMM is a method which provides for the rapid isolation of picogram quantities of DNA from targeted chromosome regions. CMM allows the rapid construction of molecular probes from chromosome regions associated with cancers. CMM therefore provides a new experimental approach to analyze specific chromosome regions associated with human diseases with significant morbidity and mortality. d. Technical improvements for genetic analysis Provide technical expertise and facilities for implementation of new techniques as they are developed (e.g., comparative genomic hybridization). The exciting technological advancement known as comparative genomic hybridization (Kallioniemi et al.,1 992) produces a map of DNA sequence copy number as a function of chromosomal location throughout the entire genetic genome. Test DNA (from cancer cells) and reference DNA (from normal cells) are differentially labeled and hybridized simultaneously to normal human peripheral blood chromosome metaphase spreads. This hybridization is detected with two different fluorochromes. Deletions, duplications or amplifications are observed as changes in the ration of the intensities of the two fluorochromes along the chromosome.

细胞遗传学实验室共享资源的目的是提供 细胞遗传学分析，基因映射和染色体显微解剖 圣安东尼奥癌症研究所成员的服务。技术 细胞遗传学分析所需的专业知识和设备，基因映射 和染色体显微解剖使其成本不佳且不良 独立研究人员自己进行这些研究 实验室。癌症中心成员的过去选择已经去过 商业上这些服务的合同或与临床合作 与患者样本分析有关的服务实验室。 该核心的目的是提供圣安东尼奥癌 研究所成员负担得起的综合细胞遗传学研究 研究标本的服务实际成本。 一个。肿瘤标本和细胞系的细胞遗传学分析 细胞系或肿瘤标本的染色体分析：技术 将提供包括G伴侣（胰蛋白酶，赖特的污渍，硼酸盐），q- 带，c-和r带，G-11，高分辨率带。 10-15 将分析中期扩散，并产生核型。这 实验室的建立用于分析各种组织的染色体 类型包括：组织培养的细胞（粘附或悬浮细胞） 外周血细胞，骨髓和实体瘤。染色体分析 还将执行人体性的体细胞杂化细胞。 b。染色体原位杂交 提供的技术包括（a）鱼（荧光原位杂交） 分析染色体标记或用宇宙映射的基因映射和（b） 放射性标记的原位杂交与小插入物进行基因映射。 c。染色体显微解剖和微量成分（CMM） 染色体显微解剖，用于特定的分子分析 将提供染色体区域。这个最近描述了技术 允许从中取出的小块染色质 染色体。此方法为起点的克隆提供了克隆 来自特定染色体区域的分子分析 疾病（Rohme等，1984； Ludecke等，1989）。有许多 与肿瘤相关的特定染色体区域 此过程可能对新遗传的产生非常宝贵 肿瘤标记。 CMM是一种提供快速隔离的方法 来自靶向染色体区域的DNA的皮克图数量。 CMM允许 染色体区域的分子探针的快速构造 与癌症相关。 CMM因此提供了新的实验 分析与人相关的特定染色体区域的方法 具有明显发病率和死亡率的疾病。 d。遗传分析的技术改进 提供技术专长和设施来实施新的 开发的技术（例如，比较基因组 杂交）。令人兴奋的技术进步称为 比较基因组杂交（Kallioniemi等，1 992）产生A DNA序列拷贝数的地图作为染色体位置的函数 在整个遗传基因组中。测试DNA（来自癌细胞）和 参考DNA（来自正常细胞）被差异标记，并且 同时杂交与正常的人类外周血染色体 中期扩散。该杂交用两个不同的 荧光色素。删除，重复或放大被认为 沿着两个荧光色素强度的分量变化 染色体。