Construction of an Unmarked Zymomonas mobilis Mutant Using a Site-Specific FLP Recombinase

Flippase expression was carried out in Zymomonas mobilis strain ZM4. The FRT-flanked selection marker gene was first integrated into the ZM4 chromosome by homologous recombination. The Saccharomyces cerevisiae flp gene was then introduced under the control of the ZM4 gap gene promoter (Pgap, encodin...

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Main Authors:	Shao-Lan Zou, Jie-Fang Hong, Cui Wang, Xin Jing, Min-Hua Zhang
Format:	Article
Language:	English
Published:	University of Zagreb 2012-01-01
Series:	Food Technology and Biotechnology
Subjects:	Zymomonas mobilis homologous recombination flippase expression transformation unmarked mutants
Online Access:	http://hrcak.srce.hr/file/139047

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spelling	doaj-195848745c3f440389d59f7a94805ed12020-11-25T02:36:58ZengUniversity of ZagrebFood Technology and Biotechnology1330-98621334-26062012-01-01504406411Construction of an Unmarked Zymomonas mobilis Mutant Using a Site-Specific FLP RecombinaseShao-Lan Zou0Jie-Fang Hong1Cui Wang2Xin Jing3Min-Hua Zhang4Biomass Conversion Laboratory, Tianjin R&D Center For Petrochemical Technology, Tianjin University, Tianjin, 300072, P.O. Box 6888, PR ChinaBiomass Conversion Laboratory, Tianjin R&D Center For Petrochemical Technology, Tianjin University, Tianjin, 300072, P.O. Box 6888, PR ChinaBiomass Conversion Laboratory, Tianjin R&D Center For Petrochemical Technology, Tianjin University, Tianjin, 300072, P.O. Box 6888, PR ChinaBiomass Conversion Laboratory, Tianjin R&D Center For Petrochemical Technology, Tianjin University, Tianjin, 300072, P.O. Box 6888, PR ChinaState Key Laboratory of Engines, Tianjin University, Tianjin, 300072, P.O. Box 6888, PR ChinaFlippase expression was carried out in Zymomonas mobilis strain ZM4. The FRT-flanked selection marker gene was first integrated into the ZM4 chromosome by homologous recombination. The Saccharomyces cerevisiae flp gene was then introduced under the control of the ZM4 gap gene promoter (Pgap, encoding glyceraldehyde-3-phosphate dehydrogenase) or the λ bacteriophage cI857-pR contained in the broad-host-range cloning vector pBBR1-MCS-2. This study demonstrated that flp was expressed and that the deletion frequency of the FRT-flanked marker gene was very high (approx. 100 %). In addition, the flp gene expression vector could be conveniently removed from the resulting unmarked Z. mobilis mutants by serially transferring the cells three times into antibiotic-free medium, thereby establishing an efficient method for constructing unmarked Z. mobilis mutants.http://hrcak.srce.hr/file/139047Zymomonas mobilishomologous recombinationflippase expressiontransformationunmarked mutants
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Shao-Lan Zou Jie-Fang Hong Cui Wang Xin Jing Min-Hua Zhang
spellingShingle	Shao-Lan Zou Jie-Fang Hong Cui Wang Xin Jing Min-Hua Zhang Construction of an Unmarked Zymomonas mobilis Mutant Using a Site-Specific FLP Recombinase Food Technology and Biotechnology Zymomonas mobilis homologous recombination flippase expression transformation unmarked mutants
author_facet	Shao-Lan Zou Jie-Fang Hong Cui Wang Xin Jing Min-Hua Zhang
author_sort	Shao-Lan Zou
title	Construction of an Unmarked Zymomonas mobilis Mutant Using a Site-Specific FLP Recombinase
title_short	Construction of an Unmarked Zymomonas mobilis Mutant Using a Site-Specific FLP Recombinase
title_full	Construction of an Unmarked Zymomonas mobilis Mutant Using a Site-Specific FLP Recombinase
title_fullStr	Construction of an Unmarked Zymomonas mobilis Mutant Using a Site-Specific FLP Recombinase
title_full_unstemmed	Construction of an Unmarked Zymomonas mobilis Mutant Using a Site-Specific FLP Recombinase
title_sort	construction of an unmarked zymomonas mobilis mutant using a site-specific flp recombinase
publisher	University of Zagreb
series	Food Technology and Biotechnology
issn	1330-9862 1334-2606
publishDate	2012-01-01
description	Flippase expression was carried out in Zymomonas mobilis strain ZM4. The FRT-flanked selection marker gene was first integrated into the ZM4 chromosome by homologous recombination. The Saccharomyces cerevisiae flp gene was then introduced under the control of the ZM4 gap gene promoter (Pgap, encoding glyceraldehyde-3-phosphate dehydrogenase) or the λ bacteriophage cI857-pR contained in the broad-host-range cloning vector pBBR1-MCS-2. This study demonstrated that flp was expressed and that the deletion frequency of the FRT-flanked marker gene was very high (approx. 100 %). In addition, the flp gene expression vector could be conveniently removed from the resulting unmarked Z. mobilis mutants by serially transferring the cells three times into antibiotic-free medium, thereby establishing an efficient method for constructing unmarked Z. mobilis mutants.
topic	Zymomonas mobilis homologous recombination flippase expression transformation unmarked mutants
url	http://hrcak.srce.hr/file/139047
work_keys_str_mv	AT shaolanzou constructionofanunmarkedzymomonasmobilismutantusingasitespecificflprecombinase AT jiefanghong constructionofanunmarkedzymomonasmobilismutantusingasitespecificflprecombinase AT cuiwang constructionofanunmarkedzymomonasmobilismutantusingasitespecificflprecombinase AT xinjing constructionofanunmarkedzymomonasmobilismutantusingasitespecificflprecombinase AT minhuazhang constructionofanunmarkedzymomonasmobilismutantusingasitespecificflprecombinase
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Construction of an Unmarked Zymomonas mobilis Mutant Using a Site-Specific FLP Recombinase

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