Replacement of the macronuclear ribosomal RNA genes of a mutant Tetrahymena using electroporation

Gene
Eduardo OriasE H Blackburn

Abstract

The macronucleus of the ciliate Tetrahymena contains approx. 10(4) ribosomal RNA gene molecules (rDNA) in the form of linear, autonomously replicating palindromes. Previous studies have shown that macronuclear rDNA molecules derived from wild-type (wt) inbred strain C3 out-replicate those derived from wt inbred strain B, in macronuclei initially heterozygous for both, leading to the complete loss of the B rDNA. However, rmm-1, a cis-acting laboratory-induced mutation obtained previously by mutagenesis of inbred strain C3, causes the mutant rmm-1 rDNA to be completely out-replicated by B rDNA. These findings suggest the following hierarchy of replication potential: wt C3 greater than wt B greater than C3-rmm-1. We used electroporation to test whether cells containing only rmm-1 macronuclear rDNA are favorable recipients for transformation with either wt B or C3 donor rDNA molecules. The donor rDNA molecules carried the selectable marker Pmr (paromomycin resistance) located in the coding region of the 17S rRNA. Transformants were obtained, at a frequency greater than 1 in 10(5), by electroporation under a wide range of electrical discharge parameters. The fraction of cells surviving electroporation varied between 2 and greater th...Continue Reading

References

Jul 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·G L YuE H Blackburn
Jul 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·A LøvlieE Orias
May 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·P J BrunsE H Blackburn
Jun 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·M M Tondravi, M C Yao
Jul 31, 1987·Cell·D D LarsonE H Blackburn
Aug 11, 1983·Nucleic Acids Research·E H BlackburnC C Johnson

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Citations

Jul 7, 1999·Gene·E PiccinniA Kraev
Nov 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·G L Yu, E H Blackburn
Nov 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·M C Yao, C H Yao
Oct 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·J Gaertig, M A Gorovsky
May 1, 1991·The Journal of protozoology·D D LarsonW L Shaiu
Apr 3, 1999·The Journal of Eukaryotic Microbiology·A J BoileauY Saimi

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