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Genetic improvement of xylanase.

Yuan Zhao. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 89-96). / Abstracts in English and Chinese. / Abstract (English) --- p.i / Abstract (Chinese) --- p.iii / Acknowledgements --- p.iv / Declaration --- p.v / Abbreviations --- p.vi / Table of Contents --- p.viii / List of Tables --- p.xii / List of Figures --- p.xiii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Lignocelluloses --- p.2 / Chapter 1.1.1 --- component of lignocellulose --- p.2 / Chapter 1.1.2 --- Xylans --- p.3 / Chapter 1.2 --- Degradation of lignocellulose --- p.9 / Chapter 1.3 --- "Endo-β-1,4- xylanase" --- p.12 / Chapter 1.3.1 --- Structure of xylanase --- p.12 / Chapter 1.3.2 --- Mode of action --- p.17 / Chapter 1.3.3 --- Appications of xylanase --- p.20 / Chapter 1.4 --- Aims of my study --- p.24 / Chapter Chapter 2 --- Materials and Methods --- p.25 / Chapter 2.1 --- Cloning of xylanase genes --- p.26 / Chapter 2.1.1 --- Materials --- p.26 / Chapter 2.1.1.1 --- Bacterial and fungal strains --- p.26 / Chapter 2.1.1.2 --- Growth media --- p.26 / Chapter 2.1.1.3 --- Vector --- p.26 / Chapter 2.1.1.4 --- Reagents for agarose gel electrophoresis --- p.27 / Chapter 2.1.1.5 --- Reagents for preparation of competent cells --- p.27 / Chapter 2.1.2 --- Methods --- p.29 / Chapter 2.1.2.1 --- Isolation of chromosomal DNA --- p.29 / Chapter 2.1.2.2 --- Amplification of exons of xylanase genes --- p.29 / Chapter 2.1.2.3 --- Agarose gel electrophoresis of DNA --- p.37 / Chapter 2.1.2.4 --- DNA recovery from agarose gel --- p.37 / Chapter 2.1.2.5 --- Assemble and amplify the full length genes --- p.38 / Chapter 2.1.2.6 --- Restriction endonuclease digestion --- p.39 / Chapter 2.1.2.7 --- Ligation of purified DNA fragment into vector --- p.39 / Chapter 2.1.2.8 --- Transformation --- p.40 / Chapter 2.1.2.9 --- Methods for making competent cells --- p.40 / Chapter 2.1.2.10 --- Plasmid DNA preparation --- p.40 / Chapter 2.1.2.11 --- DNA sequencing --- p.41 / Chapter 2.2 --- Mutagenesis of xylanase --- p.43 / Chapter 2.2.1 --- Amplification of xylanases genes --- p.47 / Chapter 2.2.2 --- DNA random mutagenesis --- p.48 / Chapter 2.2.2.1 --- DNase digestion --- p.48 / Chapter 2.2.2.2 --- Reassembly of DNA fragments --- p.48 / Chapter 2.2.2.3 --- Amplification of full-length genes --- p.48 / Chapter 2.2.2.4 --- Construction of library --- p.49 / Chapter 2.2.3 --- Screening of mutants --- p.49 / Chapter 2.2.3.1 --- Preparation of RBB-xylan --- p.49 / Chapter 2.2.3.2 --- Plate assay for screening of mutants --- p.50 / Chapter 2.3 --- Expression of xylanase genes --- p.51 / Chapter 2.4 --- Enzyme assays --- p.52 / Chapter 2.4.1 --- Xylanase assay with RBB-xylan --- p.52 / Chapter 2.4.2 --- Xylanase assay with DNS-method --- p.52 / Chapter 2.4.2.1 --- Reagents --- p.53 / Chapter 2.4.2.2 --- Xylose standard curve --- p.53 / Chapter 2.4.2. 3 --- Activity assay --- p.54 / Chapter 2.4.2. 4 --- Thermostability assay --- p.54 / Chapter Chapter 3 --- Results --- p.55 / Chapter 3.1 --- Cloning of xylanase genes --- p.56 / Chapter 3.2 --- Mutagenesis of xylanase --- p.59 / Chapter 3.2.1 --- DNA random mutagenesis --- p.59 / Chapter 3.2.2 --- Screening of mutants --- p.67 / Chapter 3.3 --- Enzyme assays --- p.69 / Chapter Chapter 4 --- Discussions --- p.76 / Chapter 4.1 --- Gene shuffling --- p.77 / Chapter 4.2 --- Screening method and activity assay --- p.78 / Chapter 4.3 --- Sequence analysis --- p.80 / Chapter 4.4 --- Future work --- p.88 / Bibliography --- p.89

Identiferoai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_324908
Date January 2004
ContributorsZhao, Yuan., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
Source SetsThe Chinese University of Hong Kong
LanguageEnglish, Chinese
Detected LanguageEnglish
TypeText, bibliography
Formatprint, xiv, 96 leaves : ill. ; 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/)

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