scholarly journals Highly Efficient Method for Introducing Successive Multiple Scarless Gene Deletions and Markerless Gene Insertions into the Yersinia pestis Chromosome

2008 ◽  
Vol 74 (13) ◽  
pp. 4241-4245 ◽  
Author(s):  
Wei Sun ◽  
Shifeng Wang ◽  
Roy Curtiss
Keyword(s):  
Yersinia Pestis ◽  
Efficient Method ◽  
Genetic Modification ◽  
Gene Deletion ◽  
Gene Deletions ◽  
Live Attenuated Vaccines ◽  
Highly Efficient ◽  
Λ Red ◽  
Red Recombination

ABSTRACT An efficient two-step recombination method for markerless gene deletion and insertion that can be used for repetitive genetic modification in Yersinia pestis was developed. The method combines λ Red recombination and counterselective screening (sacB gene) and can be used for genetic modification of Y. pestis to construct live attenuated vaccines.

scholarly journals High-Efficiency Scarless Genetic Modification in Escherichia coli by Using Lambda Red Recombination and I-SceI Cleavage

2014 ◽  
Vol 80 (13) ◽  
pp. 3826-3834 ◽  
Author(s):  
Junjie Yang ◽  
Bingbing Sun ◽  
He Huang ◽  
Yu Jiang ◽  
Liuyang Diao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Genetic Modification ◽  
Target Gene ◽  
Donor Plasmid ◽  
Content Type ◽  
Recognition Sites ◽  
Genetic Modifications ◽  
Pcr Targeting ◽  
Λ Red ◽  
Red Recombination

ABSTRACTGenetic modifications of bacterial chromosomes are important for both fundamental and applied research. In this study, we developed an efficient, easy-to-use system for genetic modification of theEscherichia colichromosome, a two-plasmid method involving lambda Red (λ-Red) recombination and I-SceI cleavage. An intermediate strain is generated by integration of a resistance marker gene(s) and I-SceI recognition sites in or near the target gene locus, using λ-Red PCR targeting. The intermediate strain is transformed with a donor plasmid carrying the target gene fragment with the desired modification flanked by I-SceI recognition sites, together with a bifunctional helper plasmid for λ-Red recombination and I-SceI endonuclease. I-SceI cleavage of the chromosome and the donor plasmid allows λ-Red recombination between chromosomal breaks and linear double-stranded DNA from the donor plasmid. Genetic modifications are introduced into the chromosome, and the placement of the I-SceI sites determines the nature of the recombination and the modification. This method was successfully used forcadAknockout,gdhAknock-in, seamless deletion ofpepD, site-directed mutagenesis of the essentialmetKgene, and replacement ofmetKwith theRickettsiaS-adenosylmethionine transporter gene. This effective method can be used with both essential and nonessential gene modifications and will benefit basic and applied genetic research.

A convenient and highly efficient method for the protection of aldehydes using very low loading hydrous ruthenium(III) trichloride as catalyst

2004 ◽  
Vol 45 (41) ◽  
pp. 7719-7721 ◽  
Author(s):  
Jian-Ying Qi ◽  
Jian-Xin Ji ◽  
Chi-Hung Yueng ◽  
Hoi-Lun Kwong ◽  
Albert S.C. Chan
Keyword(s):  
Efficient Method ◽  
Highly Efficient

Efficient synthesis of π-extended phenazasilines for optical and electronic applications

2014 ◽  
Vol 50 (99) ◽  
pp. 15760-15763 ◽  
Author(s):  
Huanhuan Li ◽  
Yang Wang ◽  
Kai Yuan ◽  
Ye Tao ◽  
Runfeng Chen ◽  
...  
Keyword(s):  
Organic Electronics ◽  
Efficient Method ◽  
Optoelectronic Properties ◽  
Efficient Synthesis ◽  
Highly Efficient ◽  
Device Applications

The rhodium-catalyzed synthesis of phenazasilines from readily achievable biarylhydrosilanes is presented. This highly efficient method offers opportunities for preparing π-extended phenazasilines with enhanced optoelectronic properties for device applications in organic electronics.

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