...toxic effects of CcdB. Another commercially available, CcdB-resistant strain is ccdB Survival™ from InvitrogenTM... the popular GatewayTM method possible: ccdB. But what is ccdB, what role does it play in modern cloning...harnessed it to evolve cloning technology. CcdB: A potent toxin... The ccdB gene, located on the F sex factor ...maintenance during cell division. ccdB codes for the toxic protein (CcdB) that acts as a DNA gyrase poison...toxic CcdB. Cells that lose ccdA through the loss of the F plasmid, succumb to the toxicity of CcdB.   ... this system contains ccdB. A successful insertion will completely replace ccdB with the investigator’...the “swapping out” of ccdB with the investigator’s gene of interest.   CcdB resistant E. coli strains...

... of CcdB. Applications of ccdB in plasmid cloning Perhaps the most widely used application of CcdB is ...the ccdB region (for “coupled cell division”). The encoded toxin-antitoxin system consists of CcdB (toxin...when the ccdB gene is replaced by the gene of interest. As a result, cells that retain the ccdB gene die...toxin) and CcdA (antitoxin) where CcdB inferes with DNA replication by binding the GyrA subunit of DNA...is GatewayTM cloning, where the ccdB gene serves as a counterselectable marker in the destination vector...a higher percentage of transformants contain the ccdB-less construct. Other toxin-antitoxin systems and...and their applications MazE/MazF Aside from CcdB/CcdA, there are many protein-based toxin-antitoxin systems...

... a E. coli strain sensitive to CcdB (e.g. DH5α, TOP10, Mach1). The ccdB gene is present in the donor vectors... BP or LR reactions. Since the CcdB protein inhibits the growth of CcdB sensitive E. coli strains, most... clone and a byproduct containing the ccdB gene. Since the ccdB product is toxic to the cell, Gateway ... attL sites. As a byproduct of the reaction, the ccdB gene is excised from the donor vector.   ...in the BP reaction, a DNA fragment containing the ccdB gene is excised from the destination vector.  Once... construct. Read our recent Plasmids 101 Post on CcdB for more information. How to clone using Gateway...efficiency The positive (antibiotic) and negative (CcdB) selection markers used for Gateway Cloning can ...

...uses the toxin CcdB. Gateway cloning takes advantage of this property by including ccdB on its destination...destination plasmids. In the destination plasmid, the ccdB gene is flanked by recombination sites. To introduce...interest at these sites in the plasmid excises the ccdB gene and the cells will only grow if they contain...lost the sacB gene. SacB can be used similarly to CcdB. For example, in the gene expression plasmid pGTvL1...

...cloning. Lentiviral. Jorge Ferrer 87067 pcDNA3.1-ccdB-Nanoluc NanoLuc® Creation of N-terminal Nanoluc ...using Gateway cloning Mikko Taipale 87075 pLenti6.2-ccdB-Nanoluc NanoLuc® Creation of C-terminal Nanoluc ... Lentival. Mikko Taipale 87078 pLenti6.2-Nanoluc-ccdB NanoLuc® Creation of N-terminal Nanoluc fusions ... co-expressed. Oskar Laur 87070 pcDNA3.1-Nanoluc-ccdB NanoLuc® Creation of N-terminal Nanoluc fusions ...using Gateway cloning Mikko Taipale 87065 pcDNA3.1-ccdB-Renilla Renilla Creation of C-terminal Renilla luciferase...using Gateway cloning Mikko Taipale 87066 pcDNA3.1-ccdB-Firefly Firefly Creation of C-terminal Firefly luciferase...Gateway cloning Mikko Taipale 87068 pcDNA3.1-Renilla-ccdB Renilla Creation of N-terminal Renilla luciferase...

...systems, including Xenopus pAG423GPD-ccdB-HA - C-terminal 3xHA tag for yeast...expression with N-terminal GFP pAG306GAL-ccdB - Yeast Gateway expression vector... vector (TEF1 promoter, CEN/ARS) pAG304GAL-ccdB - Yeast Gateway expression vector... vector (PGK1 promoter, 2μ) pAG305GAL-ccdB - Yeast Gateway expression vector... vector (PGK1 promoter, 2μ) pAG303GAL-ccdB - Yeast Gateway expression vector...

...Genotype ccdB Survival 2 T1R   TOP10 derivative. For propagating plasmids expressing the ccdB gene (important...of protein synthesis Ptrc-ccdA   Propagation of ccdB-containing plasmids Hte   High transformation ...derivative. For propagating plasmids expressing the ccdB gene (important in Gateway cloning). F- gyrA462 ...

... plasmids containing the ccdB gene, such as Gateway vectors, choose the ccdB Survival strain. For plasmids...should propagate your plasmids in: DH5α, NEB Stable, ccdB Survival, Pir1, or other. Whenever possible, we ...

...plasmids containing the ccdB gene, such as Gateway vectors, we recommend the ccdB Survival strain. Figure...propagate your plasmids in: DH5α, NEB Stable, or ccdB Survival.­ Whenever possible, we propagate plasmids...

... the ccdB gene containing BsaI restriction enzyme sites at its ends to generate pEcgRNA. The ccdB serves...

...coli strains, including DH5alpha, XL1 Blue, TOP10, ccdB survivalTM, and Stbl3TM. How to transform your plasmid...

...plasmid containing the lethal gene. An example is the ccdB gene system commonly used in Gateway® Cloning. There...

...a single insert), mostly due to the use of toxic ccdB to prevent any non-recombinant vectors from propagating...

...coli fertility plasmid that also encodes a toxin, CcdB, and the resistance to that toxin, CcdA. Knowing...

...strain DH5a, but others may require strains such as ccdB survival (for Gateway plasmids).  We also like to...

...and distribution at Addgene are DH5α, NEB stable, ccdb survival, and Pir1. The majority of plasmids contain...

...60244 Mammalian Gibson none S. pyogenes Zack pHL-H1-ccdB-mEF1a-RiH 60601 Mammalian BamHI/EcoRI none S. pyogenes...53121 Mammalian/Lentiviral BsmBI none S. pyogenes ccdB Wei pLKO5.sgRNA.EFS.PAC 57825 Mammalian/Lentiviral...

...contains a Gateway cassette containing the ccdB gene, a ccdB survival strain is necessary. What cell line...

...the desired vector. The pCoofy vectors contain the ccdB cassette, which is not copied during the PCR amplification...

...England BioLabs E. coli B F dcm ompT hsdS(rB mB) gal ccdB Survival Invitrogen F- mcrA Delta(mrr-hsdRMS-mcrBC...