MOLECULAR GENETICS OF PARAMECIUM

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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.