by Wing-Hoi Tam. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 95-99). / Abstracts in English and Chinese. / Title --- p.1 / Acknowledgement --- p.2 / Contents --- p.3 / Abbreviations --- p.5 / List of figures --- p.7 / List of tables --- p.9 / Primers and their sequences used in the projcet --- p.10 / Abstract --- p.12 / Introduction / Small heat shock proteins --- p.16 / Human heart sequencing EST project --- p.17 / Small heat shock protein B3 (HSPB3) --- p.18 / The alpha crystallins --- p.18 / Point mutation of a conserved arginine in the alpha-crystallin domain perturbs the overall alpha crystallins structure and function --- p.19 / HSP27 Confers thermotolerance in cells --- p.19 / Chaperone properties of sHSP --- p.20 / In Viro-chaperone function of sHSPs --- p.21 / Does the alpha-crystallin domain necessary for chaperone activity? --- p.21 / Expression HSPB3 in heart diseases patients and in porcine model --- p.22 / Small heat shock proteins in development and differentiation --- p.23 / HSPB3 role in mammalian embryogenesis --- p.24 / Structure of the small heat shock proteins --- p.25 / Crystal structure of a small heat shock protein --- p.25 / Subunit contacts --- p.26 / Muscle is the only tissue that simultaneously expresses all five sHSPs --- p.26 / "Two independent sHSP systems in muscle cells, to serve and to protect" --- p.27 / Methods and Materials / Small scale preparation of plasmid DNA --- p.29 / Large scale preparation of plasmid DNA --- p.29 / Amplification of mouse and rat HSPB3 coding region --- p.30 / Northern Blotting and Hybridisation --- p.31 / Purification of DNA fragments by GENECLEAN --- p.31 / Preparation of competent Escherichia coli for transformation --- p.32 / Transformation of DNA plasmid into competent Escherichia coli --- p.32 / 5´ة Rapid Amplification of cDNA Ends (5´ةRACE) --- p.33 / 3´ة Rapid Amplification of cDNA Ends (3´ةRACE) --- p.35 / Upstream and Downstream Genomic sequence --- p.36 / Isolation of genomic DNA from human tissue --- p.37 / Multiple tissue cDNA panel (MTC) --- p.37 / The yeast two hybrid screenings and confirmation --- p.43 / Isolation of total RNA from human tissues --- p.43 / Automated Sequencing --- p.44 / Alignment of DNA and amino acid sequence --- p.44 / Radiation hybrid mapping --- p.45 / Results / Determination of the foll length of human HSPB3 cDNA by 5' and 3' RACE --- p.46 / Identification of human HSPB3 gene in mouse and rat heart tissues --- p.49 / "Sequence homology between human, mouse and rat HSPB3" --- p.51 / Chromosomal mapping of the HSPB3 gene --- p.60 / Expression of human HSPB3 is not confined to the heart and skeletal muscle --- p.62 / Mouse HSPB3 expression is confined to the heart tissue --- p.65 / HSPB3 genomic PCR of downstream sequence --- p.67 / HSPB3 upstream genomic sequence --- p.67 / No intronic sequences between the open reading frame --- p.68 / The yeast two hybrid screening --- p.71 / HSPB3 can not form dimers --- p.82 / Expression of Prepro-CDD-ANF in human adult and fetal --- p.83 / Discussions / Previously reported sequence HSPL27 is a chimera --- p.87 / HSPB3 monomers do not dimerises --- p.88 / No intronic sequence in the HSPB3 gene --- p.89 / HSPB3 and CDD-ANF may play an important role during stress to the heart --- p.91 / Change of expression pattern of Prepro-CDD-ANF during development --- p.93 / Reference --- p.95
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_323096 |
| Date | January 2000 |
| Contributors | Tam, Wing-Hoi., Chinese University of Hong Kong Graduate School. Division of Biochemistry. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, 99 leaves : ill. ; 30 cm. |
| Rights | Use 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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