Auto-excision of selectable marker genes from transgenic tobacco via a stress inducible FLP/FRT site-specific recombination system

2009 ◽  
Vol 18 (3) ◽  
pp. 455-465 ◽  
Author(s):  
Hee-Jong Woo ◽  
Hyun-Suk Cho ◽  
Sun-Hyung Lim ◽  
Kong-Sik Shin ◽  
Si-Myung Lee ◽  
...  
Keyword(s):  
Transgenic Tobacco ◽  
Selectable Marker ◽  
Marker Genes ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Selectable Marker Genes ◽  
Recombination System ◽  
Site Specific Recombination System

Heat shock induced excision of selectable marker genes in transgenic banana by the Cre-lox site-specific recombination system

2012 ◽  
Vol 159 (4) ◽  
pp. 265-273 ◽  
Author(s):  
Borys Chong-Pérez ◽  
Rafael G. Kosky ◽  
Maritza Reyes ◽  
Luis Rojas ◽  
Bárbara Ocaña ◽  
...  
Keyword(s):  
Heat Shock ◽  
Selectable Marker ◽  
Marker Genes ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Selectable Marker Genes ◽  
Recombination System ◽  
Transgenic Banana ◽  
Site Specific Recombination System

Construction of Marker-Free GFP Transgenic Tobacco by Cre/lox Site-Specific Recombination System

2008 ◽  
Vol 7 (9) ◽  
pp. 1061-1070 ◽  
Author(s):  
Hong-yuan SONG ◽  
Xue-song REN ◽  
Jun SI ◽  
Cheng-qiong LI ◽  
Ming SONG
Keyword(s):  
Transgenic Tobacco ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Recombination System ◽  
Marker Free ◽  
Site Specific Recombination System

Generation of selectable marker-free transgenic eggplant resistant to Alternaria solani using the R/RS site-specific recombination system

2013 ◽  
Vol 33 (3) ◽  
pp. 411-421 ◽  
Author(s):  
Nader Ahmed Darwish ◽  
Raham Sher Khan ◽  
Valentine Otang Ntui ◽  
Ikuo Nakamura ◽  
Masahiro Mii
Keyword(s):  
Selectable Marker ◽  
Alternaria Solani ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Recombination System ◽  
Marker Free ◽  
Site Specific Recombination System

scholarly journals Marker-free genome engineering in Amycolatopsis using the pSAM2 site-specific recombination system

2021 ◽  
Author(s):  
Luísa D. F. Santos ◽  
Laëtitia Caraty-Philippe ◽  
Emmanuelle Darbon ◽  
Jean-Luc Pernodet
Keyword(s):  
Genome Engineering ◽  
High Efficiency ◽  
Genetic Manipulation ◽  
Marker Genes ◽  
Biotechnological Applications ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Recombination System ◽  
Marker Free ◽  
Site Specific Recombination System

ABSTRACTActinobacteria belonging to the genus Amycolatopsis are important for antibiotic production and other valuable biotechnological applications such as biodegradation or bioconversion. Despite their industrial importance, tools and methods for the genetic manipulation of Amycolatopsis are less developed than in other actinobacteria such as Streptomyces. Moreover, most of the existing methods do not support convenient marker-free genome engineering. Here, we report the use of the pSAM2 site-specific recombination system for the efficient deletion of marker genes or large DNA regions in Amycolatopsis. For this purpose, we constructed a shuttle vector, replicating in Escherichia coli and Amycolatopsis, expressing the Xis and Int proteins from the Streptomyces integrative and conjugative element pSAM2. These proteins are sufficient for site-specific recombination between the attachment sites attL and attR. We also constructed two plasmids, replicative in E. coli but not in Amycolatopsis, for the integration of the recombination sites attL and attR on each side of a region targeted for deletion. We exemplified the use of these tools in Amycolatopsis mediterranei DSM 40773 by obtaining with high efficiency (>95%) a marker-free deletion of one single gene in the rifamycin biosynthetic gene cluster or of the entire 90-kb cluster.IMPORTANCEThe genus Amycolatopsis is regarded as an important source of diverse specialized metabolites. Members of this genus are used in industry for the production of valuable antibiotics such as rifamycins or vancomycin. Amycolatopsis spp. also present a great interest for biotechnological applications such as biodegradation or bioconversion. Despite their importance, their genetic manipulation was somehow hampered by the lack of efficient tools. Here we report the successful use of the pSAM2 site-specific recombination system to construct unmarked deletion mutants, allowing marker recycling, or to create large deletions in A. mediterranei DSM 40773. The high efficiency of this site-specific recombination system and it possible application to other Amycolatopsis species open new opportunities for marker-free genome engineering in this genus.

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