PMID: 9529100Apr 7, 1998Paper

Electrotransformation and expression of bacterial genes encoding hygromycin phosphotransferase and beta-galactosidase in the pathogenic fungus Histoplasma capsulatum

Infection and Immunity
J P WoodsW E Goldman

Abstract

We developed an efficient electrotransformation system for the pathogenic fungus Histoplasma capsulatum and used it to examine the effects of features of the transforming DNA on transformation efficiency and fate of the transforming DNA and to demonstrate fungal expression of two recombinant Escherichia coli genes, hph and lacZ. Linearized DNA and plasmids containing Histoplasma telomeric sequences showed the greatest transformation efficiencies, while the plasmid vector had no significant effect, nor did the derivation of the selectable URA5 marker (native Histoplasma gene or a heterologous Podospora anserina gene). Electrotransformation resulted in more frequent multimerization, other modification, or possibly chromosomal integration of transforming telomeric plasmids when saturating amounts of DNA were used, but this effect was not observed with smaller amounts of transforming DNA. We developed another selection system using a hygromycin B resistance marker from plasmid pAN7-1, consisting of the E. coli hph gene flanked by Aspergillus nidulans promoter and terminator sequences. Much of the heterologous fungal sequences could be removed without compromising function in H. capsulatum, allowing construction of a substantially s...Continue Reading

References

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Mar 1, 1989·Microbiological Reviews·J R Fincham
Aug 11, 1987·Nucleic Acids Research·M Budarf, E Blackburn
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Jun 1, 1996·Infection and Immunity·Y C ChangK J Kwon-Chung

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Citations

Apr 22, 2003·Infection and Immunity·P T MageeDana Davis
Jul 1, 2004·Molecular Microbiology·Chad A RappleyeWilliam E Goldman
May 19, 2007·Journal of Medical Microbiology·Robert ZarnowskiJon P Woods
Apr 12, 2005·Eukaryotic Cell·Megan L Bohse, Jon P Woods
Jun 7, 2006·Annual Review of Microbiology·Chad A Rappleye, William E Goldman
Sep 12, 2013·Protein Science : a Publication of the Protein Society·Grainne O'KeeffeSean Doyle
Nov 6, 2013·Infection and Immunity·Andrew L GarfootChad A Rappleye
Jul 21, 2005·Molecular Biology of the Cell·M Paige NittlerAnita Sil
Jan 21, 2006·Medical Mycology·Renata De B A SoaresMaria Sueli S Felipe
Oct 14, 2006·Molecular Microbiology·Christopher L MarionWilliam E Goldman
Aug 13, 2014·Infection and Immunity·Eric D HolbrookChad A Rappleye
Oct 12, 2004·Medical Mycology·Claudia V LealJuan G McEwen
Mar 6, 2007·Eukaryotic Cell·Megan L Bohse, Jon P Woods
Feb 9, 2010·Journal of Bacteriology·Amalia PortaBruno Maresca
Sep 18, 2007·Eukaryotic Cell·A George SmulianGeorge S Deepe
Oct 5, 2018·Evolution Letters·Colin S MaxwellDaniel R Matute
Feb 5, 2002·Trends in Microbiology·V Magrini, W E Goldman

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