by Chow Wing Kuen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1993. / Includes bibliographical references (leaves 94-109). / Abstract --- p.i / Acknowledgements --- p.iii / Table of contents --- p.v / List of Tables --- p.viii / List of Figures --- p.ix / Abbreviations --- p.xi / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter Chapter 2 --- Literature review / Chapter 2.1 --- Hypersensitivity to Crustacea --- p.3 / Chapter 2.2 --- Characterization of allergens of shrimp --- p.10 / Chapter 2.3 --- Cross reactivity of crustacean allergens --- p.18 / Chapter 2.4 --- Molecular cloning and expression of allergens --- p.22 / Chapter Chapter 3 --- Immunological characterization of shrimp allergens / Chapter 3.1 --- Introduction --- p.26 / Chapter 3.2 --- Material and Methods / Chapter 3.2.1 --- Animals --- p.28 / Chapter 3.2.2 --- Sera --- p.28 / Chapter 3.2.3 --- Shrimp tissue extract --- p.29 / Chapter 3.2.4 --- Dot blotting --- p.29 / Chapter 3.2.5 --- Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.30 / Chapter 3.2.6 --- Immunoblotting --- p.32 / Chapter 3.3.7 --- Immunological detection of IgE binding proteins --- p.32 / Chapter 3.3 --- Results / Chapter 3.3.1 --- Detection of allergens in raw and cooked shrimp muscle extract --- p.35 / Chapter 3.3.2 --- Detection of allergens in hepatopancreas and ovary of the shrimp --- p.38 / Chapter 3.3.3 --- Detection of allergens in boiling shrimp fluid --- p.42 / Chapter 3.3.4 --- Detection of allergens in dried shrimp --- p.48 / Chapter 3.3.5 --- Reactivity of IgE from the shrimp- sensitive subjects with extracts from different species of penaeid shrimp --- p.48 / Chapter 3.3.6 --- Reactivity of IgE from the shrimp- sensitive subjects with muscle extracts of crustaceans and mollusks --- p.52 / Chapter 3.4 --- Discussion --- p.55 / Chapter Chapter 4 --- "Construction and immunoscreening of the cDNA library from muscle of the shrimp, Metapenaeus ensis" / Chapter 4.1 --- Introduction --- p.63 / Chapter 4.2 --- Materials and Methods / Chapter 4.2.1 --- Animals --- p.66 / Chapter 4.2.2 --- Sera --- p.66 / Chapter 4.2.3 --- Controlling ribonuclease activity --- p.66 / Chapter 4.2.4 --- Isolation of total RNA --- p.67 / Chapter 4.2.5 --- Isolation of mRNA / Chapter 4.2.5.1 --- Oligo-d(T) cellulose chromatography --- p.68 / Chapter 4.2.5.2 --- Magnetic separation --- p.70 / Chapter 4.2.6 --- Synthesis of double stranded cDNA --- p.71 / Chapter 4.2.7 --- Generation of EcoRI cohesive ends on cDNA --- p.72 / Chapter 4.2.8 --- Ligation of cDNA with λgtll vector --- p.74 / Chapter 4.2.9 --- In vitro packaging --- p.74 / Chapter 4.2.10 --- Titration of phage library --- p.75 / Chapter 4.2.11 --- Absorption of anti-E.coli antibodies --- p.76 / Chapter 4.2.12 --- Immunoscreening of the shrimp muscle cDNA library --- p.77 / Chapter 4.3 --- Results --- p.80 / Chapter 4.4 --- Discussion --- p.89 / Chapter Chapter 5 --- General conclusion --- p.92 / References --- p.94