MOLECULAR GENETICS OF PARAMECIUM

草履虫的分子遗传学

基本信息

批准号：
2842227
负责人：
CHING KUNG
金额：
$ 25.81万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
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/2842227
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

项目摘要

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.

黑甲在研究几种动物现象方面具有优势，非常适合生物电性和细胞行为的遗传解剖。在过去的30年中，Ca2+电流，K+电流和Na+电流有缺陷的突变体已被分离和表征表征。我们最近开发了一种与表型相对应的系统克隆映射基因的方法。这种方法需要表型互补，并确保克隆在其生物学背景下起作用的基因。我们计划使用这种方法从上述突变体中收集基因。我们已经克隆了控制睫状Ca2+电流的PWA和PWB基因。根据谓词基因产物的特征，我们将继续测试其生化功能。这些和其他基因产物可能会为纤毛和精子尾部的信号转导和Ca2+代谢提供见解。基因也与其他生物的已知基因同源化。我们将继续找到用这些同源基因敲定的这些同源基因来创建活的甲虫突变体的方法。将尝试三种方法：基因沉默，基莫拉皮术以及内部消除序列的操纵。因为一种便利的转换方法，可以促进整个社区的研究，所以我们还计划完美地转化Paramecia en Masse。