Six of the eight methods were less sensitive than originally reported, and five of them were considered unsuitable for use in clinical microbiology practice
Mean relative fluorescence was plotted against CAT concentration to generate the following graph
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3
Chloramphenicol is an antibiotic and is in the class of antimicrobials that inhibits protein synthesis
Objective To standardize a procedure for the determination of the enzymatic assay of chloramphenicol acetyltransferase
1 Acetyl CoA - Acetyl Coenzyme A Chloramphenicol acetyltransferase assay When a transient or stable transfection assay is developed for a promoter, a primary objective is to quantify promoter strength
In this protocol, cells transfected with an Escherichia coli transposon chloramphenicol acetyltransferase (CAT) reporter plasmid are lysed by repeated cycles of freezing and thawing and cellular debris is removed by centrifugation
Chloramphenicol acetyltransferase transfers an acetyl group (−COCH 3) from acetyl coenzyme A to two possible positions on the chloramphenicol molecule
A chloramphenicol (CAP)-binding aptamer of 80 nucleotides (nt) was reported in 2011
A very popular reporter gene and protein has been chloramphenicol acetyltransferase or CAT
Among the genes regulated by AgmR is a set of 17 genes that Chloramphenicol and β-lactams, except for ceftiofur, were dissolved in sterile demineralized water, while ceftiofur was dissolved in pure methanol according to the manufacturer's instructions
The percentage conversion of substrate to acetylated product can be calculated using the following equation: % conversion = I I + I
2
An event-specific PCR assay can confirm the presence of a specific ASE, and an isoform-specific PCR assay enables the analysis of the relative expression level of a particular RNA isoform
Fluorescent chloramphenicol as a substitute for radioactive [14C]-chloramphenicol for CAT reporter assays in Plasmodium falciparum Mol Biochem Parasitol
Affiliation 1 Department of The H
• Mg 2+ played a vital role in the aptamer and chloramphenicol interaction process
2 μ-Ci), and 250 mM Tris-Cl (pH 7
) In plants, the CAT system is generally used in transient, rather than in stable, gene expression studies
The detection of antimicrobial resistance, especially against aminoglycosides and chloramphenicol
The Charm ROSA test detects chloramphenicol below the US action level of 1 ppb and below the EU Minimum Required Performance Limit (MRPL) for chloramphenicol of 0
While the butyryl CoA remains in the aqueous phase so it does not give any scintillation, the 14 C-labeled butyryl-chloramphenicol turns more hydrophobic and diffuses linearly over time into the organic phase of the The assay was used to detect chloramphenicol-susceptible isolates in Malawi, where rapid de-escalation from broad-spectrum β-lactams such as ceftriaxone could be possible
In 2014, it was truncated to 40 nt and has since been used by most researchers, although a careful binding study is still lacking
Protein-protein interactions and protein modifications play central roles in all living organisms
The 3-acetoxy chloramphenicol can then undergo a nonenzymatic rear- rangement that results in the formation of l-acetoxy chloramphenicol
Author links open overlay panel Xin Wang a 1, Jiahao Li a 1, Dan Jian a b, Yue Zhang a, Yanke Shan a, Shouyu Wang a b, Fei Liu a
However, the previously reported aptamer for chloramphenicol suffers from low affinity, probably as a result of steric hindrance due to its bulky nature (80 nucleotides) leading to lower sensitivity in analytical assays
MeSH terms The next day, 100 to 500 random colonies from this plate were patched onto both LB and LB/chloramphenicol plates to check for colonies which lost chloramphenicol resistance (indicating plasmid loss)
Sensitive assays are available for each of these proteins that facilitate detection and quantitation of transgene expression
Chloramphenicol Acetyl Transferase Assay Michael R, Davey, Nigel W
Split Chloramphenicol Acetyl-Transferase Assay Reveals Self
To standardize a procedure for the determination of the enzymatic assay of
Chloramphenicol is often formulated as the cinnamate, palmitate (1
A chloramphenicol (CAP)-binding aptamer of 80 nucleotides (nt) was reported in 2011
Figure 3 shows a comparison of
We describe an aptamer-based colorimetric assay for chloramphenicol (CAP) based on the ability of anti-single-stranded DNA antibody (anti-ssDNA Ab) to recognize ssDNA, and the catalytic ability of PowerVision (PV), which is a polymeric conjugate of horseradish peroxidase and antibody with a high enzyme-to-antibody ratio
5% salt agar are used
calidolactis spores in agar
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The American CLSI [], on the other hand, admits interchangeable use of broth and agar dilution for most bacteria and antibiotics
5-65 mg/l) and therefore is suitable for detection below and above the The colorimetric assays indicated that chloramphenicol inhibited MM cell proliferation in a dose- and time-dependent manner (Figure 1A–1B), but only weakly inhibited the proliferation of normal PBMCs (Figure (Figure1C)
Fluorescent chloramphenicol as a substitute for radioactive [14C]-chloramphenicol for CAT reporter assays in Plasmodium falciparum Mol Biochem Parasitol
Affiliation 1 Department of The H
The original chloramphenicol aptamer containing 80 bases was truncated to 30 bases with high affinity by the SYBR Green I assay
CAT is typically assayed in a 180-μl reaction mixture containing cell extracts (or intact cells) (10–150 μl), 1–10 μl [14 C]Cm (0
A microbial assay is a biological test that determines the potency of a compound by determining the amount needed to produce effects on test organism
) In plants, the CAT system is generally used in transient, rather than in stable, gene expression studies