Identification and characterization of novel FE65-interacting proteins.

• Other Authors: Cheng, Wai Hang.
• Format: Others
• Language: English; Chinese
• Published: 2009
• Subjects: Nerve tissue proteins; Nuclear proteins; Membrane proteins; G proteins; Nerve Tissue Proteins; Nuclear Proteins; Amyloid beta-Protein Precursor; ADP-Ribosylation Factors
• Online Access: http://library.cuhk.edu.hk/record=b5896580; http://repository.lib.cuhk.edu.hk/en/item/cuhk-326879

Cheng, Wai Hang. === Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. === Includes bibliographical references (leaves 76-88). === Abstract also in Chinese. === Acknowledgement --- p.i === 摘要 --- p.iii === List of Abbreviations --- p.iv === List of Figures --- p.vi === List of Tables --- p.vii === Chapter Chapter 1 --- Introduction --- p.1 === Chapter 1.1 --- FE65 --- p.1 === Chapter 1.1.1 --- FE65 Protein Family and Their Structures --- p.2 === Chapter 1.1.1.2 --- PTB domains --- p.5 === Chapter 1.1.2 --- Expression Pattern of FE65 Proteins --- p.6 === Chapter 1.1.3 --- FE65 Family-Transgenic Animals --- p.7 === Chapter 1.1.4 --- Interacting Partners of FE65 --- p.8 === Chapter 1.1.4.1 --- "APP, APLPl and APLP2" --- p.9 === Chapter 1.1.4.2 --- LRP1 and ApoEr2 --- p.10 === Chapter 1.1.4.3 --- c-Abl --- p.11 === Chapter 1.1.4.4 --- Mena and EVL --- p.11 === Chapter 1.1.4.5 --- Tip60 --- p.12 === Chapter 1.1.4.6 --- SET --- p.12 === Chapter 1.1.4.7 --- Estrogen Receptor a --- p.13 === Chapter 1.1.4.8 --- Teashirt --- p.13 === Chapter 1.1.4.9 --- CP2/LSF/LBP1 --- p.13 === Chapter 1.1.4.10 --- Dexra sl --- p.14 === Chapter 1.1.4.11 --- P2X2-receptor subunit --- p.14 === Chapter 1.1.4.12 --- Tau --- p.15 === Chapter 1.1.4.13 --- Notchl --- p.15 === Chapter 1.1.4.14 --- Alcadein --- p.16 === Chapter 1.1.4.15 --- CD95/Fas/Apo -1 ligand --- p.16 === Chapter 1.1.4.16 --- p68 subunit of pre -mRNA cleavage and polyadenylation factor Im (p68 CFIm) --- p.17 === Chapter 1.1.4.17 --- Ataxinl --- p.17 === Chapter 1.1.5.1 --- FE65 as an adaptor protein --- p.20 === Chapter 1.1.5.2 --- FE65 and Alzheimer´ةs disease --- p.20 === Chapter 1.1.5.3 --- Transcriptional / Post-transcriptional regulation --- p.22 === Chapter 1.1.5.4 --- Apoptosis and cell cycle regulation --- p.23 === Chapter 1.1.5.5 --- Neuronal positioning and cell migration --- p.23 === Chapter 1.1.5.6 --- Learning and memory --- p.25 === Chapter 1.2 --- Objectives --- p.26 === Chapter Chapter 2 --- Investigation of the interaction between FE65 and Arf6 --- p.27 === Chapter 2.1 --- Materials --- p.27 === Chapter 2.1.1 --- DNA contructs --- p.27 === Chapter 2.1.2 --- Cell culture --- p.27 === Chapter 2.1.3 --- Immunoblotting --- p.28 === Chapter 2.1.4 --- Miscellaneous --- p.28 === Chapter 2.2 --- Methods --- p.29 === Chapter 2.2.1 --- Preparation of Escherichia coli competent cells --- p.29 === Chapter 2.2.2 --- DNA preparation with Intron Plasmid DNA --- p.30 === Chapter 2.2.3 --- DNA preparation with Macherey-Nagel NucleoBond Xtra Midi --- p.30 === Chapter 2.2.4 --- DNA preparation by the alkaline lysis method --- p.31 === Chapter 2.2.5 --- Spectrophotometric analysis of DNA --- p.32 === Chapter 2.2.6 --- Agarose gel electrophoresis --- p.32 === Chapter 2.2.7 --- Cell culture and transfection --- p.33 === Chapter 2.2.8 --- Bacterial GST-pull down assay --- p.33 === Chapter 2.2.9 --- GST-pull down assay for testing direct interaction between FE65 and Arf6 --- p.34 === Chapter 2.2.10 --- Mammalian GST-pull down assay --- p.35 === Chapter 2.2.11 --- Immunoprecipitation --- p.36 === Chapter 2.2.12 --- SDS-PAGE --- p.36 === Chapter 2.2.13 --- Immunoblotting --- p.39 === Chapter 2.3 --- Results --- p.40 === Chapter 2.3.1 --- Interaction between Arf6 and FE65 --- p.40 === Chapter 2.3.2 --- Determination of the interacting domain of FE65 with Arf6 --- p.43 === Chapter 2.3.3 --- Determination if FE65 and Arf6 interact directly --- p.45 === Chapter Chapter 3 --- Production of Antisera against Arf6 and Immunostaining of FE65-Arf6 --- p.47 === Chapter 3.1 --- Materials --- p.47 === Chapter 3.1.1 --- Protein expression and purification --- p.47 === Chapter 3.1.2 --- Immunization and harvest of antisera --- p.48 === Chapter 3.1.3 --- Immunostaining --- p.48 === Chapter 3.2 --- Methods --- p.48 === Chapter 3.2.1 --- Protein expression and purification --- p.48 === Chapter 3.2.2 --- Bradford assay --- p.50 === Chapter 3.2.3 --- Immunization --- p.50 === Chapter 3.2.4 --- Antibody purification --- p.51 === Chapter 3.2.5 --- Immunostaining --- p.52 === Chapter 3.3 --- Results --- p.53 === Chapter 3.3.1 --- Recombinant Arf6 expression and purification --- p.53 === Chapter 3.3.2 --- Titering of antisera --- p.57 === Chapter 3.3.3 --- Determination of antisera specificity --- p.59 === Chapter Chapter 4 --- Discussion --- p.68 === Chapter Chapter 5 --- Future Perspectives --- p.73 === References --- p.76