pullorum S06004 to inactivate the hsdM gene, with the help of plasmid pKD46 by the k-red method, to con- These are pKD4 or pKD13 harboring the kanamycin resistance gene (aph), or pKD3 harboring the chloramphenicol resistance gene (CAT), and different priming sites upstream and downstream of the targeting cassette
Wanner , To construct pSCI, we cloned the KpnI fragment containing the chloramphenicol resistance gene (Cm R) from plasmid pKD3 and the BamHI fragment containing the sacB gene from plasmid pKO3 into the KpnI and BamHI sites of pST76-K, respectively
Cloning plasmid containing the chloramphenicol resistance gene insert
2023 Oct Overall, increasing chloramphenicol resistance led to decreasing maximal growth at all temperatures; however, the growth inhibition was greater at the lower and higher temperature than at 37 °C The chloramphenicol resistance gene (Cm R) from plasmid pKD3, with 54 bp of homologous sequences of the hsdM gene flanking the two ends of Cm R, was PCR amplified using primers hsdM-λ-F/R
Transconjugants were selected on TSB agar plates containing chloramphenicol (16 μg ml −1) and nalidixic acid (16 μg ml −1)
2003) was cloned into pMDIAI to replace the apramycin-resistance gene and a chloramphenicol-resistance template plasmid named pMDICI was obtained
Step 1: PCR amplification of FRT-flanked Chloramphenicol resistance gene We took the pKD3 plasmid which contains Chloramphenicol resistance flanked by FRT sequences and amplified this segment using primers which were partly homologous to the chloramphenicol resistance gene (P1 and P2), and partly homologous to the area around the ackA-pta genes Pseudomonas putida KT2440 is a chloramphenicol-resistant bacterium that is able to grow in the presence of this antibiotic at a concentration of up to 25 μg/ml
Use with SnapGene software or the free Viewer to visualize additional data and align other sequences
Template plasmid pWRG100 was generated using pKD3 as template in a PCR with primers pKD-for and pKD3-I-SceI-XbaI-rev
PMID: 1319720 No abstract available Multidrug-resistant isolates carrying the chloramphenicol resistance gene were obtained from aquatic farms in various locations in Korea and studied to determine the distribution and origin of the cat gene
2019;2004:49-61
doi: 10
Silvia Cardona
We're waiting on our transforants to grow and hopefully have the Briefly, the chloramphenicol resistance gene (cat) was amplified from the pWRG100 plasmid [56] using primers tapfwd and cheYrev, which imparted flanking homologous regions upstream and downstream When making gene disruptions using the templates and priming sites in Fig
By mouth or by injection into a vein, it is used to treat meningitis, plague, cholera, and typhoid fever
Briefly, the kanamycin resistance gene kan flanked by FRT sites was amplified by PCR using the template plasmid pKD4 and hybrid primers (Table 1)
pKD3: FRT-flanked cat gene (Cm R) in oriRγ replicon requiring the pir gene product: Datsenko and Wanner (2000) (Datsenko and Wanner, 2000)
Usually primer dimers occour for one or both following reasons: 1 - The sequence of the primers are complementary; 2 - The annealing Construction of the Deletion Mutant
PCR products containing chloramphenicol resistance cassettes flanked by 39 bp of homology to the 5′ and Construction of the Deletion Mutant
PCR products containing chloramphenicol resistance cassettes flanked by 39 bp of homology to the 5′ and Chloramphenicol is a medication used in the management and treatment of superficial eye infections such as bacterial conjunctivitis, and otitis externa
6 First, PCR fragment containing chloramphenicol-resistant gene (cat) cassettes with flanking regions of a target gene was amplified and then introduced into the chromosome of S
Chloramphenicol has been indeed often blamed for being a possible When making gene disruptions using the templates and priming sites in Fig
The plasmids pKD46 (ampicillin resistance) and pKD3 (chloramphenicol resistance) were cultured in Luria-Bertani broth (LB) medium at 30 °C (Hunan Keai Medical Instrument Co
The product was recovered and purified When making gene disruptions using the templates and priming sites in Fig
The mutagenic construct containing an insertion of cat within the waaL gene coding region was amplified using primers waaL-UF/waaL-DR and transformed into Chloramphenicol
It was originally introduced as a treatment for typhoid but now it is rarely used for this purpose because of the prevalence of multiple drug-resistant Salmonella typhi
4 kb and 3
28) [1] that detoxifies the antibiotic chloramphenicol and is responsible for chloramphenicol resistance in bacteria
7 Strain construction Escherichia coli strain DH5-a was mutagenized with a mini-Tn10-chloramphenicol transposon (mini-Tn10-cam) using a specialized l phage according to The chloramphenicol resistance allele was deleted as described in ref
The wzy gene was deleted by replacing the partial gene sequence with the chloramphenicol To further confirm that the white, chlorate resistant colonies represented true deletions, loss of the cat gene was determined by lack of growth on medium supplemented with chloramphenicol
The gene encodes a protein 220 amino acids long which has TTG as the translation initiation codon
Background
Author W V Shaw
We illustrate the process using the chloramphenicol resistance gene from pKD3, but the system works equally well using the kanamycin resistance gene from pKD4
pKD3 (chloramphenicol) or pKD4 (kanamycin) pKD3 = chloramhenicol resistance cassette
The chloramphenicol resistance gene ( Cm R) from plasmid pKD3, with 54 bp of
The chloramphenicol resistance gene cmlA is disseminated on transferable plasmids that confer multiple-drug resistance in swine Escherichia coli
A recent study of β-hemolytic Escherichia coli isolated from diarrheic swine found that 53% were resistant to chloramphenicol, a drug that has been
Overall, increasing chloramphenicol resistance led to decreasing maximal growth at all temperatures; however, the growth inhibition was greater at the lower and
pKD3 and pKD4 are identical except for the region between the FRT sites, so they create an identical scar that has stop codons in all six reading To enable identification of recipients containing SGI-4 we inserted a chloramphenicol resistance gene (cat) in the arsenic resistance locus of SGI-4
Out of the 134 isolates examined, 10 showed multidrug resistance to several different antibiotics including chloramphenicol (CP)
pCP20 is a plasmid that carries ampicillin and chloramphenicol resistance genes and exhibits temperature-sensitive replication and thermal induction of FLP synthesis (Cherepanov and
Briefly, the kanamycin resistance gene kan flanked by FRT sites was amplified by PCR using the template plasmid pKD4 and hybrid primers (Table 1)
This strategy targets a chloramphenicol resistance gene, highlighted by the patent analysis performed in this study as being harboured by a noteworthy part of GMM producing microbial fermentation products from the food and feed industry
, 2016) following a one-step inactivation method (Datsenko and Wanner, 2000)
Template plasmid of chloramphenicol resistant cassette for gene disruption, Amp r, Cm r: 13: were amplified using pKD13 or pKD3 as template plasmids, 13 subsequently treated with DpnI (New England Biolabs) and purified using the QIAquick PCR Purification Kit (QIAGEN)
The 4
UniProt accession number, gene names, protein names, peptide count in EGFP and PKD3ca samples, respectively, and log2-transformed label-free quantification protein ratio are shown in the table
Using progression tissue microarrays, we here demonstrate that the serine/threonine kinase protein kinase D3 (PKD3) is highly upregulated in estrogen receptor (ER)-negative (ER (-)) tumors
Chloramphenicol resistance gene (cmr) - ccdb gene (2) Collinsella stercoris prophage (2) Escherichia coli (19) Escherichia coli bl21 (2) Tetracycline resistance removed and replaced with chloramphenicol resistance cassette from pKD3