Transformation of tetrahymena thermophila with hypermethylated rRNA genes
Abstract
The extrachromosomal rRNA genes (rDNA) of Tetrahymena thermophila contain 0.4% N/sup 6/-methyladenine. C3 strain rDNA was isolated, hypermethylated in vitro, and microinjected into B strain host cells. Clonal cell lines were established, and transformants were selected on the basis of resistance to paromomycin, conferred by the injected rDNA. The effects of methylation by three enzymes which methylate the sequence 5'-NAT-3'', the dam, EcoRI, and ClaI methylases, were tested. Hypermethylation of the injected rDNA had no effect on transformation efficiency relative to mock-methylated controls. The injected C3 strain rDNA efficiently replaced host rDNA as the major constituent of the population of rDNA molecules. Hypermethylation of the injected DNA was not maintained through 20 to 25 cell generations.
- Authors:
- Publication Date:
- Research Org.:
- Dept. of Biology, Brandeis Univ., Waltham, MA (US)
- OSTI Identifier:
- 7143997
- Resource Type:
- Journal Article
- Journal Name:
- Mol. Cell. Biol.; (United States)
- Additional Journal Information:
- Journal Volume: 8:4
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; RIBOSOMAL RNA; GENES; TETRAHYMENA; GENETIC ENGINEERING; MUTANTS; CLONE CELLS; DRUGS; ENZYME ACTIVITY; IN VITRO; METHYLATION; SENSITIVITY; ANIMALS; CELL CULTURES; CHEMICAL REACTIONS; CILIATA; INVERTEBRATES; MICROORGANISMS; NUCLEIC ACIDS; ORGANIC COMPOUNDS; PROTOZOA; RNA; 550200* - Biochemistry; 550400 - Genetics
Citation Formats
Karrer, K M, and Yao, M C. Transformation of tetrahymena thermophila with hypermethylated rRNA genes. United States: N. p., 1988.
Web. doi:10.1128/MCB.8.4.1664.
Karrer, K M, & Yao, M C. Transformation of tetrahymena thermophila with hypermethylated rRNA genes. United States. https://doi.org/10.1128/MCB.8.4.1664
Karrer, K M, and Yao, M C. 1988.
"Transformation of tetrahymena thermophila with hypermethylated rRNA genes". United States. https://doi.org/10.1128/MCB.8.4.1664.
@article{osti_7143997,
title = {Transformation of tetrahymena thermophila with hypermethylated rRNA genes},
author = {Karrer, K M and Yao, M C},
abstractNote = {The extrachromosomal rRNA genes (rDNA) of Tetrahymena thermophila contain 0.4% N/sup 6/-methyladenine. C3 strain rDNA was isolated, hypermethylated in vitro, and microinjected into B strain host cells. Clonal cell lines were established, and transformants were selected on the basis of resistance to paromomycin, conferred by the injected rDNA. The effects of methylation by three enzymes which methylate the sequence 5'-NAT-3'', the dam, EcoRI, and ClaI methylases, were tested. Hypermethylation of the injected rDNA had no effect on transformation efficiency relative to mock-methylated controls. The injected C3 strain rDNA efficiently replaced host rDNA as the major constituent of the population of rDNA molecules. Hypermethylation of the injected DNA was not maintained through 20 to 25 cell generations.},
doi = {10.1128/MCB.8.4.1664},
url = {https://www.osti.gov/biblio/7143997},
journal = {Mol. Cell. Biol.; (United States)},
number = ,
volume = 8:4,
place = {United States},
year = {Fri Apr 01 00:00:00 EST 1988},
month = {Fri Apr 01 00:00:00 EST 1988}
}