Homologous Recombination in Q-Beta Rna Bacteriophage

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Description

Q-Beta phage RNAs with inactivating insertion (8 base) or deletion (17 base) mutations within their replicase genes were transfected into Escherichia coli spheroplasts containing QB replicase provided in trans by a resident plasmid. Replicase-defective (Rep~) Q3 phage produced by these spheroplasts were unable to form plaques on cells lacking this plasmid. When individual Rep~ phage were isolated and grown to high titer in cells containing plasmid derived Q3 replicase, revertant Q3 phage (Rep'), with the original mutation (insertion or deletion) repaired, were obtained at a frequency of ca. 1 x 108. RNA recombination via a "template switching" mechanism involving Q3 ... continued below

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iii, 40 leaves : ill

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Palasingam, Kampan May 1992.

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Q-Beta phage RNAs with inactivating insertion (8 base) or deletion (17 base) mutations within their replicase genes were transfected into Escherichia coli spheroplasts containing QB replicase provided in trans by a resident plasmid. Replicase-defective (Rep~) Q3 phage produced by these spheroplasts were unable to form plaques on cells lacking this plasmid. When individual Rep~ phage were isolated and grown to high titer in cells containing plasmid derived Q3 replicase, revertant Q3 phage (Rep'), with the original mutation (insertion or deletion) repaired, were obtained at a frequency of ca. 1 x 108. RNA recombination via a "template switching" mechanism involving Q3 replicase, the mutant phage genome, and the plasmid-derived replicase mRNA was shown to be the primary means by which these mutant phages reverted to wild type.

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iii, 40 leaves : ill

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  • May 1992

Added to The UNT Digital Library

  • March 9, 2015, 8:15 a.m.

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  • Aug. 25, 2017, 11:57 a.m.

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Citations, Rights, Re-Use

Palasingam, Kampan. Homologous Recombination in Q-Beta Rna Bacteriophage, thesis, May 1992; Denton, Texas. (digital.library.unt.edu/ark:/67531/metadc500683/: accessed November 21, 2017), University of North Texas Libraries, Digital Library, digital.library.unt.edu; .