MOLECULAR GENETICS OF PARAMECIUM

草履虫的分子遗传学

• 批准号: 6385297
• 负责人: CHING KUNG
• 金额: $ 24.28万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-05-15 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6385297
• 关键词: Paramecium; alleles; binding proteins; calmodulin; cell membrane; cilium /flagellum motility; electrophysiology; gene complementation; gene induction /repression; gene mutation; gene targeting; membrane channels; membrane proteins; microinjections; microorganism genetics; molecular cloning; phenotype; site directed mutagenesis

Paramecium offers advantages in the study of several animal phenomena and is well suited for the genetic dissection of bioelectricity and cell behavior. Over the last 30 years, mutants with defective Ca2+ currents, K+ currents and Na+ currents have been isolated and characterized phenotypically. We have recently developed a method to systematically clone Paramecium genes that correspond to phenotypes. This method entails phenotypic complementation and guarantees the genes cloned to be functional in their biological context. We plan to use this method to harvest the genes from the above mutants. We have cloned the pwA and pwB genes that control the ciliary Ca2+ currents. Based on features of the predicated gene products, we will continue to test their biochemical functions. These and other gene products may give insights to signal transductions and Ca2+ metabolism in cilia and sperm tail. Genes have also been cloned by their homology to known genes of other organisms. We will continue to find ways to create live Paramecium mutants with these homologous genes knocked out. Three methods will be tried: gene silencing, kimeraplasty, and the manipulation of internal eliminated sequences. Because a convenient transformation method with facilitate research of the whole community, we also plan to perfect methods of transforming paramecia en masse.

草履虫在研究几种动物现象方面具有优势，并且非常适合于生物电和细胞行为的遗传解剖。 在过去的30年中，突变体有缺陷的Ca 2+电流，K+电流和Na+电流已被分离和表征表型。 我们最近开发了一种方法，系统地克隆草履虫基因，对应的表型。 这种方法需要表型互补，并保证克隆的基因在其生物学背景下具有功能。 我们计划用这种方法从上述突变体中收获基因。我们已经克隆了控制睫状体Ca 2+电流的CCLA和CCLB基因。 根据预测基因产物的特征，我们将继续检测其生化功能。这些和其他基因产物可能会给纤毛和精子尾部的信号转导和Ca ~（2+）代谢的见解。基因也可以通过与其他生物的已知基因同源而被克隆。 我们将继续寻找方法来创造活草履虫突变体与这些同源基因敲除。将尝试三种方法：基因沉默，kimeraplasty，和内部消除序列的操作。由于变换方法方便，便于整个社会的研究，我们还计划完善参数变换方法。