ISOLATION OF THE GENE FOR CHEDIAK-HIGASHI SYNDROME

CHEDIAK-HIGASHI 综合征基因的分离

• 批准号: 3082900
• 负责人: RANDALL F HOLCOMBE
• 金额: $ 6.03万
• 依托单位: LOUISIANA STATE UNIV HSC SHREVEPORT
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-09-01 至 1994-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3082900
• 关键词: Chediak Higashi syndrome; T lymphocyte; complementary DNA; cytogenetics; disease /disorder model; gel electrophoresis; gene expression; gene therapy; genetic disorder; genetic disorder diagnosis; genetic library; genetic manipulation; genetic mapping; genetic markers; genetic transcription; homeostasis; human tissue; immunodeficiency; laboratory mouse; linkage mapping; molecular cloning; nucleic acid hybridization; nucleic acid probes; prenatal diagnosis; restriction fragment length polymorphism

Chediak-Higashi syndrome (CHS) is a rare immuno-deficiency disease with manifestations involving alterred hemostasis, an increase in the frequency of malignancies, abnormal function of lysosomes with resultant lack of "Natural Killer" activity, and an increase in a specific subset of T-lymphocytes. The gene responsible for the syndrome has not been characterized in either the human or murine system, although the chromosomal location of the gene causing the analogous syndrome in mice (bg) has been identified. The goals of this project are: 1) Generate a murine "jumping library" to identify new probes close to bg, 2) Define restriction fragment length polymorphisms (RFLP) with these probes through cross hybridization with human DNA, 3) Determine genetic linkage of the probes to the CHS gene by RFLP and family analysis, 4) analyze human and murine northern blots to identify transcripts of the putative gene and 5) sequence putative CHS genes and predict the protein product. The technology for identifying a gene by "reverse genetics" on the basis of its chromosomal location exists and should be facilitated in this instance by the presence of an animal model for which the gene has been previously mapped. Identification of the CHS gene should provide information about hemostasis, immune surveillance, lysosomal function, and the in vivo function of gamma delta T-lymphocytes. Dr. Randall Holcombe is completing training in the medical subspecialties of hematology and oncology and has already initiated a productive research program concerning CHS. He has developed expertise in the molecular biological techniques required for the successful completion of the research program described herein over the prior two years. The Department of Genetics, where the bulk of the research will be performed, has the equipment necessary for recombinant DNA research, with tissue culture and animal facilities. The Hematology Division and the Department of Genetics sponsor journal clubs, lab meetings, and several lecture series, and provide an exciting and stimulating environment for research.

Chediak-Higashi 综合征 (CHS) 是一种罕见的免疫缺陷疾病 涉及止血改变、频率增加的表现 恶性肿瘤、溶酶体功能异常导致缺乏 “自然杀手”活动，以及特定子集的增加 T淋巴细胞。导致该综合征的基因尚未被证实 其特征在于人类或小鼠系统，尽管 引起小鼠类似综合征的基因的染色体位置 (bg)已被确定。 该项目的目标是：1）生成小鼠“跳跃库” 识别接近 bg 的新探针，2) 定义限制性片段长度 通过与这些探针的交叉杂交进行多态性（RFLP） 人类 DNA，3) 通过以下方式确定探针与 CHS 基因的遗传连锁 RFLP 和家族分析，4) 分析人类和小鼠 Northern 印迹 识别推定基因的转录本和 5) 推定 CHS 序列 基因并预测蛋白质产物。 以“反向遗传学”为基础来鉴定基因的技术 它的染色体位置存在并且在这种情况下应该被促进 通过先前已经研究过该基因的动物模型的存在 映射。 CHS 基因的鉴定应提供以下信息： 止血、免疫监视、溶酶体功能和体内 γ δ T 淋巴细胞的功能。 Randall Holcombe 博士即将完成医学亚专业的培训 血液学和肿瘤学，并已经启动了富有成效的研究 有关 CHS 的计划。他在分子生物学领域积累了丰富的专业知识 成功完成该任务所需的生物技术 本文描述的前两年的研究计划。部门 大部分研究将在遗传学研究所进行， 重组 DNA 研究所需的设备，包括组织培养和 动物设施。血液科和遗传学系 赞助期刊俱乐部、实验室会议和多个系列讲座，以及 为研究提供令人兴奋和刺激的环境。