Return to search

The Beta-lactamases of ampicillin-resistant, Escherichia coli.

by Ling Kin Wah, Thomas. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1991. / Includes bibliographical references (leaves 103-117). / ABSTRACT --- p.i / ACKNOWLEDGMENTS --- p.v / LIST OF ABBREVIATIONS --- p.vi / TABLE OF CONTENTS --- p.viii / LIST OF TABLES --- p.xv / LIST OF FIGURES --- p.xix / INTRODUCTION --- p.1 / LITERATURE REVIEW --- p.2 / Chapter 1. --- Structure of the bacterial cell envelope --- p.2 / Chapter 2 . --- The β-lactam antibiotics --- p.4 / Chapter 3. --- Mode of action of β-lactam antibiotics --- p.5 / Chapter 4. --- Penicillin-binding proteins (PBPs) --- p.6 / Chapter 5. --- Mechanisms of bacterial resistance to β-lactam antibiotics --- p.7 / Chapter 5.1 --- Non-enzymatic resistance --- p.7 / Chapter 5.1.1 --- Alteration in cell permeability --- p.8 / Chapter 5.1.2 --- Alteration of the target site --- p.9 / Chapter 5.1.3 --- Tolerance and persistence --- p.9 / Chapter 5.2 --- Enzyme-mediated resistance --- p.12 / Chapter 6. --- Transfer of resistance --- p.13 / Chapter 7. --- β-lactamases --- p.16 / Chapter 7.1 --- History --- p.16 / Chapter 7.2 --- Classification of β-lactamases --- p.17 / Chapter 7.2.1 --- Richmond and Sykes scheme --- p.17 / Chapter 7.2.2 --- Matthew scheme --- p.18 / Chapter 7.2.3 --- Bush scheme --- p.19 / Chapter 7.3 --- β-lactamases of Gram-negative bacteria --- p.19 / Chapter 7.3.1 --- Chromosomally-mediated β-lactamases --- p.19 / Chapter 7.3.2 --- Plasmid-mediated β-lactamases --- p.20 / Chapter 7.4 --- β-lactamase inhibitors --- p.25 / Chapter 7.5 --- Regulation of β-lactamase production --- p.28 / Chapter 7.5.1 --- β-lactamase induction --- p.28 / Chapter 7.5.2 --- Mutation to constitutive enzyme production --- p.29 / Chapter 7.5.3 --- β-lactam induced β-lactamase production --- p.30 / Chapter 8. --- Emergence of resistance due to production of β-lactamases --- p.31 / Chapter 8.1 --- Resistance in staphylococci --- p.32 / Chapter 8.2 --- Resistance in haemophili and gonococci --- p.33 / Chapter 8.3 --- Resistance in Enterobacteriaceae (non E. coli) --- p.34 / Chapter 8.4 --- Distribution of β-lactamases in E. coli --- p.35 / MATERIALS AND METHODS / Chapter 1. --- Bacterial strains --- p.38 / Chapter 1.1 --- Standard organisms --- p.38 / Chapter 1.2 --- Clinical isolates --- p.38 / Chapter 2. --- Antibiotics --- p.39 / Chapter 3. --- "Media, chemicals and culture conditions" --- p.39 / Chapter 4. --- Bacterial identification and viable bacterial count --- p.39 / Chapter 5. --- Antibiotic sensitivity testing --- p.40 / Chapter 5.1 --- Disk diffusion --- p.40 / Chapter 5.2 --- Determination of minimal inhibitory concentration (MIC) --- p.40 / Chapter 6. --- Plasmid analysis --- p.41 / Chapter 6.1 --- Transfer of drug resistance plasmids --- p.41 / Chapter 6. 2 --- Molecular studies of plasmids --- p.42 / Chapter 6.2.1 --- Extraction of plasmid DNA --- p.43 / Chapter 6.2.2 --- Agarose gel electrophoresis --- p.43 / Chapter 6.2.3 --- Molecular size determination --- p.44 / Chapter 7 . --- DNA hybridization --- p.44 / Chapter 7.1 --- DNA blotting --- p.44 / Chapter 7.1.1 --- Colony blotting --- p.45 / Chapter 7.1.2 --- Southern blotting --- p.45 / Chapter 7.2 --- Labeling of oligonucleotide probe --- p.46 / Chapter 7.3 --- Hybridization --- p.47 / Chapter 7.4 --- Autoradiography --- p.47 / Chapter 7.5 --- Re-use of blots --- p.48 / Chapter 8. --- Detection and screening for classification of β-lactamases --- p.48 / Chapter 8.1 --- Detection of β-lactamases --- p.48 / Chapter 8.1.1 --- Acidimetric --- p.48 / Chapter 8.1.2 --- Chromogenic substrate --- p.49 / Chapter 8.1.2.1 --- Whole cell --- p.49 / Chapter 8.1.2.2 --- Cell extract and filtrate --- p.49 / Chapter 8.2 --- Screening for classification of β-lactamases --- p.49 / Chapter 9. --- "Preparation, purification, qualitative and quantitative analyses of the β-lactamase from transconjugants TU117, TB117 and the recipient K12" --- p.51 / Chapter 9.1 --- Large scale preparation of enzyme --- p.51 / Chapter 9.2 --- Gel filtration --- p.52 / Chapter 9.3 --- Preparative isoelectric focusing (PIEF) --- p.53 / Chapter 9.4 --- Protein determination --- p.55 / Chapter 9.5 --- Qualitative analyses and characterization of β-lactamases --- p.56 / Chapter 9.5.1 --- Analytical isoelectric focusing --- p.56 / Chapter 9.5.1.1 --- Semi-quantitative determination of β-lactamases --- p.56 / Chapter 9.5.1.2 --- Polyacrylamide gel preparation --- p.57 / Chapter 9.5.1.3 --- Isoelectric focusing --- p.58 / Chapter 9.5.1.4 --- pH measurement --- p.58 / Chapter 9.5.1.5 --- Gel development and recording --- p.59 / Chapter 9.5.1.5.1 --- Nitrocefin staining --- p.59 / Chapter 9.5.1.5.2 --- Silver staining --- p.59 / Chapter 9.5.1.6 --- Isoelectric point (pI) determination --- p.60 / Chapter 9.5.2 --- Spectrophotometric assay of β-lactam substrates --- p.60 / Chapter 9.5.2.1 --- Absorption spectra of β-lactam antibiotics --- p.60 / Chapter 9.5.2.2 --- The molar extinction coefficient of β-lactam substrates --- p.60 / Chapter 9.5.2.3 --- Measurement of β-lactamase hydrolytic activities --- p.61 / Chapter 9.5.2.4 --- Determination of enzyme kinetics --- p.61 / Chapter 9.5.3 --- Molecular weight determination of proteins --- p.62 / Chapter 9.5.3.1 --- SDS-polyacrylamide gel preparation --- p.62 / Chapter 9.5.3.1.1 --- Resolving gel --- p.62 / Chapter 9.5.3.1.2 --- Stacking gel --- p.63 / Chapter 9.5.3.2 --- Electrophoresis --- p.63 / Chapter 9.5.3.3 --- Staining and recording --- p.64 / Chapter 9.5.3.4 --- Molecular weight determination --- p.64 / RESULTS / Chapter 1. --- Collection of organisms --- p.65 / Chapter 2 . --- Identification of organisms --- p.65 / Chapter 3. --- Antibiotic sensitivity testing --- p.66 / Chapter 4. --- Genetic and molecular studies of ampicillin- resistant plasmids --- p.68 / Chapter 4.1 --- Transfer of ampicillin-resistant factor --- p.68 / Chapter 4.1.1 --- E. coli K12 14R525 as recipient --- p.68 / Chapter 4.1.2 --- other Enterobacteriaceae --- p.68 / Chapter 4.2 --- Plasmid studies of E. coli --- p.69 / Chapter 5. --- Detection and identification of β-lactamases --- p.69 / Chapter 5.1 --- Analytical IEF --- p.70 / Chapter 5.2 --- DNA hybridization --- p.70 / Chapter 5.2.1 --- Colony blot hybridization --- p.70 / Chapter 5.2.2 --- Southern blot hybridization --- p.71 / Chapter 6. --- Characterization of TEM-1 producing E. coli --- p.71 / Chapter 6.1 --- Susceptibility testing --- p.71 / Chapter 6.2 --- Enzyme kinetic study --- p.72 / Chapter 6.2.1 --- Absorption spectra and molar extinction coefficient of β-lactam antibiotics --- p.72 / Chapter 6.2.2 --- Comparison of the substrate profiles --- p.73 / Chapter 6.3 --- Correlation of MICs to β-lactamase activities --- p.73 / Chapter 7. --- "Isolation, quantitation and characterization of β-lactamases isolated from three E. coli strains" --- p.74 / Chapter 7.1 --- Preparation of β-lactamases --- p.75 / Chapter 7. 2 --- Purification of β-lactamases --- p.76 / Chapter 7.2.1 --- Gel-filtration chromatography --- p.76 / Chapter 7.2.2 --- Preparative isoelectric focusing --- p.77 / Chapter 7.3 --- Characterization of the purified β-lactamases --- p.78 / Chapter 7.3.1 --- Isoelectric point --- p.78 / Chapter 7.3.2 --- Molecular weight assessment --- p.79 / Chapter 7.3.3 --- Enzyme kinetic study --- p.79 / DISCUSSION / Chapter 1. --- Epidemiology of ampici11in (or amoxycillin)- resistant E. coli --- p.81 / Chapter 2. --- Distribution of β-lactamases in ampicillin- resistant E. coli --- p.84 / Chapter 3. --- Correlation between level of resistance and β-lactamase activity --- p.86 / Chapter 4. --- Plasmid-mediated TEM-1 β-lactamase --- p.89 / Chapter 4.1 --- Transfer of resistance --- p.89 / Chapter 4 .2 --- Identification of β-lactamases by DNA hybridization --- p.91 / Chapter 5. --- Mechanism of high-level resistance --- p.93 / Chapter 5.1 --- Selection of resistant strains --- p.93 / Chapter 5.2 --- β-lactamases preparation and purification --- p.95 / Chapter 5.3 --- Hyperproduction of β-lactamase --- p.97 / SUMMARY AND CONCLUSIONS --- p.102 / REFERENCES --- p.103 / APPENDICES / Chapter 1. --- TABLES --- p.118 / Chapter 2. --- FIGURES --- p.153

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_318906
Date January 1991
ContributorsLing, Kin Wah., Chinese University of Hong Kong Graduate School. Division of Clinical and Pathological Sciences.
PublisherChinese University of Hong Kong
Source SetsThe Chinese University of Hong Kong
LanguageEnglish
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xx, 187, [1] leaves : ill. (some mounted) ; 30 cm.
RightsUse of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/)

Page generated in 0.0025 seconds