PMID: 9526697Apr 4, 1998Paper

Trans-kingdom conjugation offers a powerful gene targeting tool in yeast

Genetic Analysis : Biomolecular Engineering
M Nishikawa, K Yoshida

Abstract

Gene targeting is one of the powerful techniques used to investigate eukaryotic genes. In a typical eukaryotic microbe, Saccharomyces cerevisiae yeast, we examined trans-kingdom conjugation between Escherichia coli bacterium and yeast as a gene targeting tool. Here, it is shown that trans-kingdom conjugation effectively induced gene replacement even on yeast's target loci (e.g. ura3-52 allele) which is never targeted by conventional transformation. This clearly indicates that trans-kingdom conjugation offers a very powerful gene targeting tool in yeasts. In fact, Southern hybridization analysis of transconjugants distinctly verified the accuracy in the conjugative gene replacement. The efficiency of gene replacement was about 0.4 x 10(-7) per recipient yeast. This is enough to sustain gene targeting with gene replacement by trans-kingdom conjugation. We also discuss the mechanism of conjugative gene replacement.

References

Apr 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·D H Figurski, D R Helinski
May 14, 1969·Journal of Molecular Biology·H W Boyer, D Roulland-Dussoix
Mar 1, 1984·Microbiological Reviews·N Willetts, B Wilkins
Jan 1, 1984·Molecular & General Genetics : MGG·M Rose, F Winston
Jan 1, 1983·Journal of Bacteriology·H ItoA Kimura

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Citations

Mar 27, 2013·Antonie van Leeuwenhoek·Petar Tomev Mitrikeski
Sep 15, 2005·Nucleic Acids Research·Young Geol Yoon, Michael D Koob
Apr 1, 2015·Current Microbiology·Petar Tomev Mitrikeski
Dec 31, 2010·Anatomy & Cell Biology·Young Geol YoonYoung Hyun Yoo

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