MOLECULAR MECHANISMS CONTROLLING PROGRAMMED CELL DEATH

天蛾的某些可识别的神经元和肌肉 经历快速的，发育程序性细胞死亡， 变态到成年。 这些肌肉的承诺， 简并需要新的RNA和蛋白质合成。 本 这项工作旨在确定编码“细胞死亡”特异性的基因， proteins. Poly A+ RNA将用于构建阶段特异性 在λ GT 10中的消减cDNA文库，和个体 将筛选重组体以确认它们含有 发育调控的cDNA序列。 这些克隆人将 进行测序，以确定其一级结构 推定的蛋白质产物。 将这些序列与 蛋白质数据库中的那些可能使我们能够识别 它们的功能。 似乎符合几个定义的克隆 细胞死亡基因的标准将在细菌或 抗体生产。 抗体和核酸探针都将 用于开始表征所涉及的机制， 在发育过程中调节细胞死亡的Manduca和相关的 蛾类Antheraea polyphemus。 同源的表达 细胞死亡基因也将在青蛙和鸡中进行检测， 曼杜卡的抗体和核酸探针

Certain identifiable neurons and muscles of the moth Manduca sexta undergo a rapid, developmentally programmed cell death during metamorphosis to the adult. The commitment of these muscles to degenerate requires new RNA and protein synthesis. The present work seeks to identify genes which encode "cell death" specific proteins. Poly A+ RNA will be used to construct stage-specific subtraction cDNA libraries in lambda GT10, and individual recombinants will be screened to confirm that they contain developmentally regulated cDNA sequences. These clones will then be sequenced to determine the primary structure of their presumptive protein products. Comparison of these sequences to those in protein databases may allow us to identify the nature of their function. Clones which appear to meet several defined criteria for cell death genes will be expressed in bacteria or antibody production. Both antibody and nucleic acid probes will be used to begin characterization of the mechanisms involved in regulating cell death during development in Manduca and a related moth species Antheraea polyphemus. The expression of homologous cell death genes will also be examined in frog and chicken using the antibodies and nucleic acid probes from Manduca.