PiggyBac transposon-mediated mutagenesis and application in yeast Komagataella phaffii

Biotechnology Letters
Jinxiang ZhuYuanxing Zhang

Abstract

Around one-fourth of the Komagataella phaffii genes encode hypothetical proteins with unknown functions. However, lack of powerful tools for genetic screening in K. phaffii significantly limits the functional analysis of these unknown genes. Transposon mutagenesis has been utilized as an insertional mutagenesis tool in many other organisms and would be extremely valuable if it could be applied in K. phaffii. In this study, we investigated in K. phaffii the transposition activity and efficiency of piggyBac (PB) transposon, a DNA transposon from the cabbage looper moth Trichoplusia ni through the integrated-plasmid system. We also designed a binary-plasmid system which could generate stable mutants. Finally we evaluated the quality of this mutagenesis system by a simple screening for functional genes involved in K. phaffii carbon catabolite repression. Our results demonstrate that PB-mediated mutagenesis could be a feasible and useful tool for functional gene screening in K. phaffii.

References

Dec 1, 1985·Molecular and Cellular Biology·J M CreggK R Madden
Sep 28, 2002·The Journal of Biological Chemistry·Sabire Ozcan
Mar 19, 2005·International Journal of Systematic and Evolutionary Microbiology·Cletus P Kurtzman
Mar 10, 2006·Microbiology and Molecular Biology Reviews : MMBR·George M Santangelo
Oct 24, 2007·Methods in Molecular Biology·James M Cregg
Oct 24, 2007·Methods in Molecular Biology·Laura A SchroderWilliam A Dunn
Apr 22, 2008·Current Protocols in Molecular Biology·Fred Winston
May 26, 2009·Nature Biotechnology·Kristof De SchutterNico Callewaert
Jan 6, 2011·Proceedings of the National Academy of Sciences of the United States of America·Kosuke YusaNancy L Craig
Jul 28, 2012·The International Journal of Biochemistry & Cell Biology·Andriy S PolupanovAndriy A Sibirny
Jan 26, 2013·Microbial Cell Factories·Marizela DelicBrigitte Gasser
Jul 25, 2013·Biotechnology Letters·Sasha LarsenGeoff P Lin-Cereghino
Dec 11, 2013·Scientific Reports·Wentao XuYunbo Luo
Feb 19, 2014·The Journal of Cell Biology·Taras Y NazarkoSuresh Subramani
May 24, 2014·Metabolic Engineering·Justyna NoconDiethard Mattanovich
Oct 10, 2014·Nature Biotechnology·Gary Walsh
Oct 23, 2016·Microbial Cell Factories·Wei ShenMian Zhou
Aug 2, 2017·Biotechnology Advances·Jan-Philipp SchwarzhansKarl Friehs

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BETA
PCR

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BioEdit
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