Abstract

The 11 surface immobilization antigens of stock 51 of the ciliate Paramecium tetraurelia are specified by a family of 11 unlinked genes, only one of which can be expressed at a time. Expression is determined by an interaction of cytoplasmic, genetic and environmental factors. We have devised a transformation system utilizing microinjection that makes it possible to introduce fragments of DNA containing these cloned genes into deletion mutants where they are telomerized, replicated and expressed in a normal fashion. We wish to continue our preliminary observations on copy number of these introduced fragments of DNA and investigate the existence of specific DNA sequences that are involved in replication of DNA in Paramecium. A second purpose of this project is to discover the molecular mechanisms involved in control of gene expression in the system. We plan to continue experiments designed to identify, isolate, and elucidate the functional regions of the genes involved in the expression of these antigens. Deletions produced in vitro, as well as chimaeras of genes that can be expressed at different temperatures will be used in transformation experiments to identify polymerase binding sites, sites where trans acting factors bind, and possible important regions within the coding sequences. We believe that the project will contribute to the general problems of gene replication and expression.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM031745-08
Application #
3280021
Study Section
Genetics Study Section (GEN)
Project Start
1983-04-01
Project End
1993-12-31
Budget Start
1991-01-01
Budget End
1991-12-31
Support Year
8
Fiscal Year
1991
Total Cost
Indirect Cost

  Institution

Name
Indiana University Bloomington
Department
Type
Schools of Arts and Sciences
DUNS #
006046700
City
Bloomington
State
IN
Country
United States
Zip Code
47401

  Publications

Martin, L D; Pollack, S; Preer Jr, J R et al. (1994) DNA sequence requirements for the regulation of immobilization antigen A expression in Paramecium tetraurelia. Dev Genet 15:443-51
Kim, C S; Preer Jr, J R; Polisky, B (1994) Identification of DNA segments capable of rescuing a non-mendelian mutant in paramecium. Genetics 136:1325-8
Steele, C J; Barkocy-Gallagher, G A; Preer, L B et al. (1994) Developmentally excised sequences in micronuclear DNA of Paramecium. Proc Natl Acad Sci U S A 91:2255-9
Preer Jr, J R (1993) Nonconventional genetic systems. Perspect Biol Med 36:395-419
Kim, C S; Preer Jr, J R; Polisky, B (1992) Bacteriophage lambda DNA fragments replicate in the Paramecium macronucleus: absence of active copy number control. Dev Genet 13:97-102
Preer, L B; Hamilton, G; Preer Jr, J R (1992) Micronuclear DNA from Paramecium tetraurelia: serotype 51 A gene has internally eliminated sequences. J Protozool 39:678-82
Jessop-Murray, H; Martin, L D; Gilley, D et al. (1991) Permanent rescue of a non-Mendelian mutation of Paramecium by microinjection of specific DNA sequences. Genetics 129:727-34
Rudman, B; Preer, L B; Polisky, B et al. (1991) Mutants affecting processing of DNA in macronuclear development in paramecium. Genetics 129:47-56
Gilley, D; Rudman, B M; Preer Jr, J R et al. (1990) Multilevel regulation of surface antigen gene expression in Paramecium tetraurelia. Mol Cell Biol 10:1538-44
Preer Jr, J R (1989) Update on the molecular genetics of Paramecium. J Protozool 36:182-4

Showing the most recent 10 out of 16 publications