GENETIC ANALYSIS OF MEIOTIC CENTROMERE FUNCTION

减数分裂着丝粒功能的遗传分析

基本信息

批准号：
6181092
负责人：
JOHN E TOMKIEL DEAN
金额：
$ 11.5万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-07-01 至 2002-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6181092
关键词：
Drosophilidae antibody centromere gene mutation genetic mapping male meiosis molecular cloning molecular genetics

项目摘要

DESCRIPTION: The specific aims of this proposal are to genetically identify and characterize genes required for the assembly and function of centromeres in meiosis. These studies will be carried out in male Drosophila melanogaster, a model genetic organism whose centromeres appear functionally and structurally similar to human centromeres. Mutants have been induced using transposon mutagenesis, and were screened for their effects on the transmission of a meiotically unstable dicentric chromosome (a chromosome with two centromeres). Mutations that either increased or preferentially decreased meiotic transmission of this chromosome were selected. These will be characterized genetically and cytologically to identify candidate genes for centromere structural and/or regulatory proteins. In addition, a collection of deficiency-bearing chromosomes that together delete 80 percent of the autosomal euchromatin will be tested in trans for effects on the segregation of this dicentric chromosome. This deficiency screen will identify regions of the genome that contain genes important for the stability of the dicentric. Selected mutants will mapped by genetic and molecular methods, and cloned either by transposon-based methods or by chromosomal waLking. Cloned cDNAs will be expressed in bacteria or baculovirus to produce antigen for the production of polyclonal antibodies. Additionally, the nature of the dicentric chromosome will be further studied genetically. The two centromeres of the dicentric will be detached and recovered on separate chromosomes to allow tests of the properties of each centromere individually. New dicentric chromosomes will be generated as derivatives of the original dicentric. These will be characterized genetically and cytologically to identify properties that influence the mitotic and meiotic behavior of centromeres of dicentric chromosomes.

描述：该提议的具体目的是从基因上识别并表征着丝粒的组装和功能所需的基因在减数分裂。这些研究将在雄性果蝇中进行 Melanogaster，一种模型的遗传生物，其centromeres在功能上出现在结构上类似于人类的中心粒。突变体已被诱导使用转座子诱变，并筛选其对减数分裂不稳定的二含染色体的传播（染色体带有两个centromeres）。增加或优先增加的突变选择了该染色体的减数分裂传播降低。这些会从遗传和细胞学上进行特征以识别候选基因用于丝粒结构和/或调节蛋白。另外，含有缺陷的染色体的收集共同删除80％常染色体着染色质的影响将在反式中进行测试该折叠染色体的分离。这个缺陷屏幕将识别包含基因的基因组的区域 dicentric的稳定性。选定的突变体将由遗传和分子方法，并通过基于转座子的方法克隆或通过染色体步行。克隆的cDNA将在细菌或杆状病毒产生抗原以生产多克隆抗体。此外，将进一步研究折叠染色体的性质遗传。二十座的两个centromeres将被脱离，并且在单独的染色体上恢复以允许测试每种特性单独的中心。新的少染色体将被生成原始折叠的衍生物。这些将被描述从遗传和细胞学上讲，以识别影响折叠染色体的丝粒的有丝分裂和减数分裂行为。