CHROMOSOME STRUCTURE AND GENE ARRANGEMENT

这项研究的长期目标是深入了解 染色质结构和基因表达。 中所述的具体目标 这些应用是：（1）获得有关分子的新信息 中期染色体中活性基因的结构。 中期 染色体和间期核将从同步化的细胞中分离出来。 鸡细胞 活性基因的组织，包括 甘油醛-3-磷酸脱氢酶、胸苷激酶和β-微管蛋白 将使用核酸酶研究染色体/细胞核中的基因， 含有这些基因作为探针的重组DNA分子。的 中期染色体中活性基因的纵向分布将是 使用染色体切口平移方法进行研究。 (2)探讨 核骨架结构（联会复合体）在前期阶段 减数分裂细胞 抗联会复合体蛋白的单克隆抗体 将制备并用作探针，以研究结构和 核内联会复合体的生理作用。 (3)的 异常染色体中的染色质结构，如均匀 染色区域（HSR）和双分钟（dm），这是最常见的 观察耐药突变体和肿瘤细胞，将进行研究 类似地使用重组DNA和核酸酶作为探针。 的可能性 基因扩增可能发生在长春新碱耐药突变体中， 其中包含HSR和dm将使用微分 构建的cDNA文库或基因组DNA文库的筛选方法， 变种人 从这些描述的项目中获得的结果可以 提供分子遗传学方面的有用信息， 人类疾病的知识。

The long-term objectives of this research are to gain insights into chromatin structure and gene expression. The specific aims described in this application are: (1) To gain new information concerning the molecular architecture of active genes in metaphase chromosomes. Metaphase chromosomes and interphase nuclei will be isolated from synchronized chicken cells. The organizations of active genes, including the glyceradehyde-3-phosphate dehydrogenase, thymidine kinase and Beta-tubulin genes, in chromosomes/nuclei will be investigated using nucleases and recombinant DNA molecules containing these genes as probes. The longtitudinal distribution of active genes in metaphase chromosomes will be studied using a chromosome nick-translation method. (2) To investigate the nuclear skeletal structure (synaptonemal complex) in prophase stage of meiotic cells. Monoclonal antibodies against synaptonemal complex proteins will be prepared and used as probes to investigate the structural and physiological roles of the synaptonemal complex inside the nuclei. (3) The chromatin structure in the abnormal chromosomes such as homogeneously staining regions (HSR) and double minutes (dm), which are most often observed in drug-resistant mutants and neoplastic cells, will be studied similarly using recombinant DNA and nucleases as probes. The possibilities that gene amplification may take place in the vincristine-resistant mutants which contain HSR and dm will be investigated using a differential screening method of a cDNA library or genomic DNA library constructed from the mutants. The results obtained from these described projects may provide useful information in molecular genetics and enhance fundamental knowledge in human diseases.