MOLECULAR MECHANISMS CONTROLLING PROGRAMMED CELL DEATH

六纹夜蛾某些可识别的神经元和肌肉 在发育过程中经历快速的、程序性的细胞死亡 向成年变态。这些肌肉的承诺是 退化需要新的RNA和蛋白质合成。现在 这项工作旨在识别编码“细胞死亡”特异性的基因 蛋白质。Poly A+RNA将用于构建阶段特异性 在lambda GT10和个体中消减cDNA文库 将对重组子进行筛选，以确认它们含有 发育调节的cdna序列。然后这些克隆人就会 进行测序以确定它们的一级结构 推定的蛋白质产品。将这些序列与 蛋白质数据库中的那些可能会让我们识别出 它们的功能。似乎符合多个定义的克隆 细胞死亡基因的标准将在细菌或 抗体的产生。抗体和核酸探针都会 被用来开始描述 调控Manduca发育过程中的细胞死亡及其相关基因 蛾种Antheraea Polyphemus。同源基因的表达 细胞死亡基因也将在青蛙和鸡身上进行检测 Manduca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.