Yeung Kai-shing, Elvis. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 98-104). / Abstracts in English and Chinese. / Abstract (English) --- p.i / Abstract (Chinese) --- p.iii / Acknowledgement --- p.v / Table of Contents --- p.vii / List of Tables --- p.x / List of Figures --- p.xi / Abbreviations --- p.xiii / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter Chapter 2 --- Literature Review / Chapter 2.1 --- Allergy to shellfish --- p.3 / Chapter 2.2 --- Identification and characterization of crustacean allergens --- p.7 / Chapter 2.3 --- Identification and characterization of mollusk allergens --- p.10 / Chapter 2.4 --- Cross-reactivity between shellfish allergens --- p.14 / Chapter 2.5 --- Epitopes of shellfish allergens --- p.17 / Chapter Chapter 3 --- Isolation and characterization of allergens in cephalopods / Chapter 3.1 --- Introduction --- p.22 / Chapter 3.2 --- Materials and methods --- p.25 / Chapter 3.2.1 --- Animals --- p.25 / Chapter 3.2.2 --- Sera --- p.25 / Chapter 3.2.3 --- Cephalopod tissue extracts --- p.25 / Chapter 3.2.4 --- Quantitation of protein --- p.27 / Chapter 3.2.5 --- Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.27 / Chapter 3.2.6 --- Immunoblotting --- p.29 / Chapter 3.2.7 --- Immunological detection of IgE binding protein --- p.30 / Chapter 3.2.8 --- Controlling of ribonuclease (RNase) activity --- p.31 / Chapter 3.2.9 --- Isolation of total RNA --- p.31 / Chapter 3.2.10 --- Construction of cDNA library --- p.32 / Chapter 3.2.10.1 --- Synthesis of first-strand cDNA --- p.33 / Chapter 3.2.10.2 --- Amplification of cDNA by LD-PCR --- p.33 / Chapter 3.2.10.3 --- Polishing of ds cDNA --- p.34 / Chapter 3.2.10.4 --- Ligation of adaptor --- p.34 / Chapter 3.2.10.5 --- Phosphorylation of adaptor-ligated cDNA --- p.35 / Chapter 3.2.10.6 --- Size fractionation of cDNA --- p.35 / Chapter 3.2.10.7 --- Ligation of cDNA to vector --- p.36 / Chapter 3.2.10.8 --- Packaging of plasmids to bacteriophage --- p.36 / Chapter 3.2.10.9 --- Titration of the packaging reaction --- p.38 / Chapter 3.2.10.10 --- Amplification of the cDNA library --- p.39 / Chapter 3.2.11 --- Immunoscreening of octopus cDNA library --- p.40 / Chapter 3.2.12 --- Extraction of phage DNA --- p.42 / Chapter 3.2.13 --- Purification of cDNA insert from phage plasmid --- p.42 / Chapter 3.2.14 --- cDNA cloning and protein expression --- p.43 / Chapter 3.2.15 --- Nucleotide sequencing analysis of DNA --- p.45 / Chapter 3.2.16 --- Immunoblot analysis of the recombinant allergen --- p.46 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Immunoblotting of allergens in cephalopod --- p.47 / Chapter 3.3.2 --- RNA from octopus --- p.47 / Chapter 3.3.3 --- The cDNA library of octopus --- p.51 / Chapter 3.3.4 --- Immunoscreening of cDNA library from octopus --- p.56 / Chapter 3.3.5 --- Nucleotide sequencing determination and analysis --- p.61 / Chapter 3.3.6 --- Protein expression of Oct l I --- p.76 / Chapter 3.3.7 --- Immunoblot analysis of recombinant tropomyosin from octopus --- p.83 / Chapter 3.4 --- Discussion --- p.87 / Chapter Chapter 4 --- Conclusions --- p.96 / References --- p.98