Immunological and molecular studies on allergens in cephalopods.

• Other Authors: Yeung, Kai-shing Elvis.
• Format: Others
• Language: English; Chinese
• Published: 2000
• Subjects: Allergens > isolation & purification; Allergens > immunology; Allergens > genetics; Tropomyosin > immunology; Tropomyosin > genetics; Mollusca > immunology; Mollusca > genetics; Molecular Biology
• Online Access: http://library.cuhk.edu.hk/record=b5890329; http://repository.lib.cuhk.edu.hk/en/item/cuhk-323198

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