Thesis (Ph.D.)--Hong Kong University of Science and Technology, 2006. / Includes bibliographical references (leaves 195-210). Also available in electronic version.

Understanding the binding specificity of transcription factors is an important step towards accurate computational prediction of regulatory sequences governing gene expression. Higher-throughput binding site characterization methods have long been available in the laboratory fort he study of protein-DNA interactions in solution or upon a surface. In this thesis a new method is introduced for characterization of inducible regulatory sequences in living cells based on construction and analysis of promoter-reporter gene plasmids. Spiked oligonucleotides are used to generate libraries of regulatory sequences with subtle variations from a known regulatory element. Screening of the library in cell culture for the capacity of the mutated sequences to mediate expression provides a diverse collection of responsive and non-responsive sequences to aid in understanding the sequence requirement for an inducible transcription factor binding site. We apply the methodology to the study of antioxidant responsive elements, the target sites of the Nfe212 transcription factor. These target sequences commonly found in the promoters of detoxification genes modulate gene expression in response to a diverse array of chemicals. The variants serve as a primary screen for future targeted mutational analysis to further characterize context-specific sequence requirement in the ARE and/or interdependence between positions. Moreover, a transcription factor binding profile can be generated from functional ARE variants in the library screen. Such an ARE profile performs as well as standard profiles based on bona fide ARE sequences drawn from the scientific literature.

We first identified the transcriptional regulatory element of the human dynamin IV gene (Hdyn IV; dymple). The Hdyn IV belongs to a large GTPase family. This protein has a N-terminal highly conserved tripartite GTP-binding domain, coiled-coil (CC) region, but it lacks the pleckstrin homology (PH) domain and a modestly conserved C-terminal proline rich domain (PRD). Hdyn IV gene is enriched in subcellular membrane fractions of cytoplasmic vesicles and endoplasmic reticulum, and the function of Hdyn IV gene is considered to be associated with the functions of mitochondria. The Hdyn IV is expressed as four alternative splicing variants in all eukaryotic organisms. Our question concerning why expressions of four alternative splicing variants in brain tumor tissues? To elucidate the regulatory mechanism and the transcription factors involved, we firstly determined the transcriptional start site by 5¡¦ RACE. We next cloned the 5¡¦-flanking region of the Hdyn IV gene and determined the nucleotide sequence of 999 bases upstream from the transcription start site. The promoter has several potential binding sites for AP2, Sp1 binding protein, but it lacks TATA and CAAT boxes. Transfection studies using a series of Hdyn IV promoter luciferase constructs in HeLa cell demonstrate that the 5¡¦flanking region has a promoter activity. Functional promoter element of the Hdyn IV gene was located within the ¡V140~ +29 region. Deletion analyses demonstrated that the minimal promoter activity for the transcriptional element of Hdyn IV was detected in the sequence between nucleotides ¡V110 and ¡V100. Electorphoretic mobility shift assay demonstrated that a putative transcriptional factor bound to the ¡V119 to ¡V90 region. Site-directed mutagenesis analysis of this region revealed that nucleotides at positions ¡V108 to ¡V100 were essential for transactivation mediated by this element. To summary, the data indicated that the ¡¦¡¦CTCCCAGCA¡¦¡¦ (-108~ -100) sequence is capable of regulating Hdyn IV gene expression. However, the protein involved in the binding of this novel sequence requires further study.

The noroviruses are important pathogen of epidemic and sporadic gastroenteritis in all age group and show great genetic diversity. The aim of the present study was to describe the prevalence and genetic diversity of noroviruses among children hospitalized with acute sporadic gastroenteritis in Kaohsiung, Taiwan. Fecal samples were collected from hospitalized pediatric patients with sporadic gastroenteritis below age of 18 years during a 2-year period (2007 to 2008). Norovirus RNA was detected by reverse transcription-polymerase chain reaction and comfirmed by sequence analysis. Two different sets of primers were used. Region C primers target shell domain at 5¡¦ end of capsid gene and region D primers target highly variable P2 subdomain at 3¡¦ end of capsid gene. Noroviruses were identified in 5 of 39 (12%) rotavirus-negative specimens using region C primers. Using region D primers only one among these 5 samples could yield PCR product, which showed concordant noroviral genotype. 3 (10%, n=30) specimens from children below age of 5 years tested positive. All these 5 patients had symptoms of vomiting and 3 had fever. All PCR products were sequenced and showed 2 strains of genogroup 1 (G 1.4) and 3 strains of genogroup2 (G 2.4). To our knowledge, this is the first report that demostrated G1.4 genotype norovirus from Taiwan. Norovirus accounted for 10% of sporadic non-bacterial gastroenteritis cases among hospitalized children below 5 years of age in Kaohsiung, Taiwan.

Thèse de doctorat : Sciences de la vie et de la santé : Lille 1 : 2001. / Textes en français et en anglais. Résumé en français. Bibliogr. f. 59-77. Notes bibliogr.

Thèse de doctorat : Aspects moléculaires et cellulaires de la biologie : Strasbourg 1 : 2007. / Titre provenant de l'écran-titre. Bibliogr. p. 62-89.

Thesis (Ph. D.)--Hong Kong University of Science and Technology, 2002. / Includes bibliographical references (leaves 175-187). Also available in electronic version. Access restricted to campus users.

Thesis (M. Phil.)--University of Hong Kong, 2009. / Includes bibliographical references (leaves 155-169). Also available in print.

Thesis (M. Sc.)--York University,1998. Graduate Program in Biology. / Typescript. Includes bibliographical references (leaves 76-90). Also available on the Internet. MODE OF ACCESS via web browser by entering the following URL: http://wwwlib.umi.com/cr/yorku/fullcit?pMQ27358.

Thesis (Ph. D.)--University of Texas at Austin, 2001. / Vita. Includes bibliographical references (leaves 133-166). Available also in a digital version from Dissertation Abstracts.