NDLI: A mini-Mu transposon-based method for multiple DNA fragment integration into bacterial genomes

Content Provider	Springer Nature Link
Author	Wei, Xiao Xing Shi, Zhen Yu Li, Zheng Jun Cai, Lei Wu, Qiong Chen, Guo Qiang
Copyright Year	2010
Abstract	A method for construction of bacterial strains with multiple DNA inserted into their chromosomes has been developed based on the mini-Mu transposon and FLP/FRT recombination. Exogenous DNA can be integrated by Mu transposition with an FRT cassette containing selection marker and conditional replicative origin (R6Kγori). Subsequently, with the introduction of a helper plasmid bearing gene of FLP recombinase, drug-resistant selection marker is excised from the chromosome. Cells cured of the helper plasmid can undergo the next cycle of transposition and excision of selection marker. Each cycle can add further foreign gene(s) to the chromosome. As an example, resistance genes of chloramphenicol, tetracycline, and gentamicin were successively integrated into the chromosome of Escherichia coli BW25113 by three cycles of insertion and excision as described above. This method proved to be simple and time-saving, which could be applicable to a variety of microorganisms.
Starting Page	1533
Ending Page	1541
Page Count	9
File Format	PDF
ISSN	01757598
Journal	Applied Microbiology and Biotechnology
Volume Number	87
Issue Number	4
e-ISSN	14320614
Language	English
Publisher	Springer-Verlag
Publisher Date	2010-05-28
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	DNA integration Mu transposon Bacterial genome FLP/FRT recombination Transposition Microbial Genetics and Genomics Microbiology Biotechnology
Content Type	Text
Resource Type	Article
Subject	Medicine Applied Microbiology and Biotechnology Biotechnology

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