Thesis (M.S.)--Virginia Polytechnic Institute and State University, 1993. / Vita. Abstract. Includes bibliographical references (leaves 57-64). Also available via the Internet.

by Sham Lok To. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 143-171). / Abstracts in English and Chinese. / Abstract --- p.i / 摘要 --- p.iii / Acknowledgement --- p.iv / Abbreviations --- p.v / Table of contents --- p.vi / List of figures --- p.ix / List of tables --- p.xi / Chapter Chapter One --- Literature Review --- p.1 / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.1.1 --- About L. edodes --- p.3 / Chapter 1.1.2 --- Nutritional and medicinal values of L. edodes --- p.4 / Chapter 1.1.3 --- Life cycle of L. edodes --- p.6 / Chapter 1.1.4 --- Environmental factors affecting fruiting body formation in L. edodes --- p.6 / Chapter 1.2 --- Molecular mechanisms of fruiting body development in L. edodes --- p.8 / Chapter 1.2.1 --- Expression profiling and identification of differentially expressed genes during fruiting --- p.8 / Chapter 1.2.2 --- Changing in membrane structure --- p.11 / Chapter 1.2.3 --- The signal transduction cascade --- p.12 / Chapter 1.3 --- Transformation in L. edodes and in other fungi --- p.14 / Chapter 1.3.1 --- Transformation of L. edodes --- p.14 / Chapter 1.3.2 --- Transformation in other fungi --- p.17 / Chapter 1.4 --- Bioinformatics tools for comparative promoter analysis --- p.22 / Chapter 1.5 --- Objectives and significance --- p.26 / Chapter Chapter Two --- Promoter analysis of differentially expressed genes (DEGs) in the fruiting body development in L. edodes --- p.27 / Chapter 2.1 --- Introduction --- p.27 / Chapter 2.2 --- Materials and methods --- p.29 / Chapter 2.2.1 --- Strains and cultivation conditions --- p.29 / Chapter 2.2.2 --- Genome walking of the 5' flanking region of the DEGs --- p.29 / Chapter 2.2.3 --- Annealing Control Primed (ACP) PCR --- p.31 / Chapter 2.2.4 --- Construction of genomic DNA library --- p.36 / Chapter 2.2.5 --- Nested PCR to amplify the target sequences --- p.37 / Chapter 2.2.6 --- Cloning and sequencing of the 5' flanking region --- p.38 / Chapter 2.2.7 --- Determination of transcription start site by the Neural Network algorithm --- p.39 / Chapter 2.2.8 --- Identification of putative transcription factor binding sites --- p.40 / Chapter 2.3 --- Results --- p.41 / Chapter 2.3.1 --- Construction of adaptor linked template for genome walking --- p.41 / Chapter 2.3.2 --- Sequence analysis and quality control --- p.41 / Chapter 2.3.3 --- Comparison of various methods in genome walking --- p.42 / Chapter 2.3.4 --- Promoter analysis --- p.42 / Chapter 2.4 --- Discussion --- p.58 / Chapter Chapter Three --- In-silico analysis of transcription factor binding sites and identification transcription factors expressed in L. edodes --- p.64 / Chapter 3.1 --- Introduction --- p.64 / Chapter 3.2 --- Material and methods --- p.67 / Chapter 3.2.1 --- Sequence manipulation and extraction of homologous ESTs from C. cinereus --- p.67 / Chapter 3.2.2 --- Extraction of 5' flanking region of the corresponding ESTs and promoter prediction --- p.67 / Chapter 3.2.3 --- Positional cloning of mating type factor A --- p.68 / Chapter 3.3 --- Results --- p.70 / Chapter 3.3.1 --- Sequence extraction and manipulation --- p.70 / Chapter 3.3.2 --- In-silico analysis of transcription factor binding sites in C. cinereus . --- p.70 / Chapter 3.3.3 --- Comparison of putative TFBS between L. edodes and C. cinereus --- p.71 / Chapter 3.3.4 --- Identification of transcription factors in L. edodes by positional cloning --- p.71 / Chapter 3.4 --- Discussion --- p.85 / Chapter Chapter Four --- Identification，expression profiling and promoter analysis of hydrophobin genes --- p.91 / Chapter 4.1 --- Introduction --- p.91 / Chapter 4.2 --- Material and methods --- p.92 / Chapter 4.2.1 --- Clustering and grouping of the hydrophobin ESTs --- p.92 / Chapter 4.2.2 --- Identification of the consensus sequences of the hydrophobin groups --- p.93 / Chapter 4.2.3 --- RNA Sources and Preparation --- p.93 / Chapter 4.2.4 --- Expression profiling of hydrophobin genes by RT-PCR --- p.95 / Chapter 4.2.5 --- Promoter cloning and analysis of hydrophobin genes --- p.95 / Chapter 4.3 --- Results --- p.97 / Chapter 4.3.1 --- Isolation and characterization of four newly found hydrophobin genes --- p.97 / Chapter 4.3.2 --- Expression levels of hydrophobins --- p.100 / Chapter 4.3.3 --- Promoter sequencing of the hydrophobins --- p.103 / Chapter 4.4 --- Discussion --- p.103 / Chapter Chapter Five --- Transformation of L. edodes --- p.110 / Chapter 5.1 --- Introduction --- p.110 / Chapter 5.2 --- Materials and methods --- p.112 / Chapter 5.2.1 --- Vectors and primers design --- p.112 / Chapter 5.2.2 --- Maxi-preparation of plasmids --- p.112 / Chapter 5.2.3 --- Cultural condition and optimization of protoplasts release --- p.114 / Chapter 5.2.4 --- PEG mediated transformation --- p.115 / Chapter 5.2.5 --- Electroporation mediated transformation --- p.116 / Chapter 5.2.6 --- PCR screening of regenerated transformant --- p.116 / Chapter 5.2.7 --- Particle bombardment --- p.117 / Chapter 5.3 --- Results --- p.121 / Chapter 5.4 --- Discussion --- p.128 / Chapter Chapter Six --- General discussions --- p.132 / References --- p.143

Leung, Chiu Yi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 219-235). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.iii / ABSTRACT --- p.v / 摘要 --- p.vii / TABLE OF CONTENTS --- p.ix / LIST OF FIGURES --- p.xiv / LIST OF TABLES --- p.xviii / LIST OF ABBREVIATIONS --- p.xix / Chapter CHAPTER 1. --- GENERAL INTRODUCTION --- p.1 / Chapter CHAPTER 2. --- LITERATURE REVIEW --- p.5 / Chapter 2.1 --- Introduction to carotenoids --- p.5 / Chapter 2.1.1 --- Structures and general chemical properties --- p.6 / Chapter 2.1.2 --- Dietary source of carotenoids --- p.8 / Chapter 2.2 --- Biosynthesis of carotenoids in plants --- p.8 / Chapter 2.2.1 --- Formation of isopretenyl diphosphate (IPP) --- p.12 / Chapter 2.2.2 --- "Formation of C40 backbone, phytoene" --- p.13 / Chapter 2.2.3 --- Desaturation reactions --- p.16 / Chapter 2.2.4 --- Isomerization --- p.18 / Chapter 2.2.5 --- Cyclization --- p.18 / Chapter 2.2.6 --- Xanthophylls synthesis --- p.19 / Chapter 2.2.6.1 --- Hydroxylation --- p.19 / Chapter 2.2.6.2 --- Epoxidation and de-epoxidation --- p.19 / Chapter 2.2.6.3 --- Neoxanthin formation --- p.20 / Chapter 2.2.7 --- Carotenoids catabolism by cleavage enzymes --- p.20 / Chapter 2.2.8 --- Carotenoids sequestration --- p.20 / Chapter 2.2.9 --- Regulations of Carotenogenesis in plant --- p.21 / Chapter 2.3 --- Roles of carotenoids in plants --- p.24 / Chapter 2.3.1 --- Precursor of abscisic acid (ABA) production --- p.24 / Chapter 2.3.2 --- Photomorphogenesis: Prolamella body formation --- p.27 / Chapter 2.3.3 --- "Light-harvesting, energy transfer and photoprotection" --- p.30 / Chapter 2.4 --- Importance of carotenoids to human --- p.34 / Chapter 2.4.1 --- Provitamin A activity and conversion --- p.34 / Chapter 2.4.2 --- Roles of vitamin A and carotenoids in diseases prevention --- p.36 / Chapter 2.4.2.1 --- Visual cycle and related diseases --- p.36 / Chapter 2.4.2.2 --- Cardiovascular diseases --- p.37 / Chapter 2.4.2.3 --- Cancer --- p.38 / Chapter 2.4.3 --- Roles of vitamin A in gene regulation --- p.39 / Chapter 2.4.4 --- Bioavailability and daily intake recommendation --- p.39 / Chapter 2.5 --- Vitamin A deficiency (VAD) --- p.42 / Chapter 2.5.1 --- Clinicopathological features --- p.44 / Chapter 2.5.1.1 --- Visual problems --- p.44 / Chapter 2.5.1.2 --- Infection --- p.45 / Chapter 2.6 --- Global efforts in combating VAD --- p.45 / Chapter 2.6.1 --- Dietary diversification --- p.46 / Chapter 2.6.2 --- Supplementation --- p.48 / Chapter 2.6.3 --- Pro-vitamin A enriched crops by genetical engineering --- p.49 / Chapter 2.6.3.1 --- Tomato --- p.49 / Chapter 2.6.3.2 --- Potato --- p.50 / Chapter 2.6.3.3 --- Canola --- p.50 / Chapter 2.6.3.4 --- The Golden Rice (GR) project --- p.51 / Chapter 2.6.3.4.1 --- The 1st generation (GR1) --- p.52 / Chapter 2.6.3.4.2 --- The 2nd generation (GR2) --- p.54 / Chapter 2.7 --- Rice phytoene synthase as a GR candidate enzyme --- p.55 / Chapter 2.7.1 --- General properties of phytoene synthase in higher plant --- p.55 / Chapter 2.7.1.1 --- Gene duplication and structure --- p.55 / Chapter 2.7.1.2 --- Membrane association and cation requirement --- p.57 / Chapter 2.7.1.3 --- Expression pattern and tissue-specificity --- p.58 / Chapter 2.7.2 --- "Rice phytoene synthases: Ospsy1, Ospsy2 and Ospsy3" --- p.60 / Chapter 2.7.2.1 --- Gene duplication and structure --- p.60 / Chapter 2.7.2.2 --- Protein structures and phylogenetic analysis --- p.60 / Chapter 2.7.2.3 --- Expression pattern --- p.62 / Chapter 2.7.2.4 --- Light- and stress-induced expression --- p.63 / Chapter 2.7.3 --- Rice phytoene synthases activity in rice seeds --- p.64 / Chapter 2.7.3.1 --- Previous study in rice seed carotenogenic capacity --- p.64 / Chapter 2.8 --- Seed-specific rice promoters --- p.66 / Chapter 2.8.1 --- Previous studies on seed-specific expression in rice --- p.66 / Chapter 2.8.1.1 --- Endosperm-specific cis-acting regulatory elements --- p.67 / Chapter 2.8.1.2 --- Requirements to confer endosperm-specific expression in rice --- p.69 / Chapter 2.9 --- Project overview and hypothesis --- p.71 / Chapter CHAPTER 3. --- MATERIALS AND METHODS --- p.73 / Chapter 3.1 --- Chemicals --- p.73 / Chapter 3.2 --- Vectors and bacterial strains in regular cloning --- p.73 / Chapter 3.3 --- Plant materials --- p.74 / Chapter 3.4 --- Construction of gene cassettes for plant transformation --- p.74 / Chapter 3.4.1 --- Construction of gene cassettes for Ospsyl and Ospsy2 study --- p.74 / Chapter 3.4.1.1 --- Cloning of Ospsyl and Ospsy2 from rice --- p.76 / Chapter 4.1.1.1 --- Total RNA extraction --- p.75 / Chapter 3.4.1.1.2 --- Amplification of cDNA of Ospsyl by RT-PCR --- p.77 / Chapter 3.4.1.1.3 --- Amplification of cDNA of Ospsy2 by PCR --- p.80 / Chapter 3.4.1.2 --- Cloning of constitutive CaMV35S promoter --- p.82 / Chapter 3.4.1.2.1 --- Preparation of pBI121 vector --- p.82 / Chapter 3.4.1.2.2 --- Amplification of CaMV35S promoter by PCR --- p.82 / Chapter 3.4.1.3 --- Cloning of endosperm-specific rice glutelin-1 (Gt-1) promoter --- p.84 / Chapter 3. 4.1.3.1 --- Genomic DNA extraction --- p.84 / Chapter 3.4.1.3.2 --- Amplification of Gt-1 promoter by PCR --- p.84 / Chapter 3.4.1.4 --- Construction of gene cassettes for Ospsyl and Ospsy2 driven by CaMV35S and rice Gt-1 promoter --- p.87 / Chapter 3.4.2 --- Construction of gene cassettes for promoter analysis --- p.90 / Chapter 3.4.2.1 --- Cloning of full length and fragments of Ospsyl promoter --- p.92 / Chapter 3.4.2.1.1 --- Genomic DNA extraction --- p.95 / Chapter 3.4.2.1.2 --- Amplification of full length and fragments of Ospsy1 promoter --- p.95 / Chapter 3.4.2.2 --- Cloning of CaMV35S minimal promoter --- p.96 / Chapter 3.4.2.2.1 --- Amplification of CaMV35S minimal promoter --- p.97 / Chapter 3.4.2.3 --- Cloning of rice Gt-1 promoter --- p.97 / Chapter 3.4.2.3.1 --- Genomic DNA extraction --- p.97 / Chapter 3.4.2.3.2 --- Amplification of Gt-1 promoter --- p.97 / Chapter 3.4.2.4 --- Annealing of linker --- p.93 / Chapter 3.4.3.5 --- Making of rice GCN4 multimers --- p.98 / Chapter 3.4.2.6 --- Construction of gene cassettes for promoter analysis --- p.99 / Chapter 3.4.2.7 --- Construction of gene cassettes for Gt-1 promoter analysis --- p.104 / Chapter 3.4.3 --- Confirmation of sequence fidelity --- p.106 / Chapter 3.5 --- Rice transformation --- p.107 / Chapter 3.5.1 --- Plant materials --- p.107 / Chapter 3.5.2 --- Preparation of Agrobacterium --- p.107 / Chapter 3.5.3 --- Agrobacterium mediated transformation --- p.108 / Chapter 3.5.4 --- Callus induction from mature rice seeds --- p.109 / Chapter 3.5.6 --- Co-cultivation and selection --- p.109 / Chapter 3.5.7 --- Pre-regeneration and regeneration of transgenic rice --- p.110 / Chapter 3.5.8 --- Plantation of transgenic rice --- p.110 / Chapter 3.6 --- Detection of transgene expression --- p.112 / Chapter 3.6.1 --- Detection at DNA level --- p.112 / Chapter 3.6.1.1 --- Genomic DNA extraction --- p.112 / Chapter 3.6.1.2 --- PCR screening --- p.122 / Chapter 3.6.1.3 --- Synthesis of DIG-labeled DNA probes --- p.116 / Chapter 3.6.1.4 --- Southern blot analysis --- p.118 / Chapter 3.6.2 --- Detection at RNA level --- p.119 / Chapter 3.6.2.1 --- Total RNA extraction --- p.119 / Chapter 3.6.2.2 --- Northern blot analysis --- p.119 / Chapter 3.6.3 --- Detection at protein level --- p.119 / Chapter 3.6.3.1 --- Antibody production --- p.119 / Chapter 3.6.3.1 --- Ospsyl and Ospsy2 induction in pET system --- p.120 / Chapter 3.63.1.2 --- Immunization of rabbit and serum collection --- p.123 / Chapter 3.6.3.2 --- Total protein extraction from plant materials --- p.124 / Chapter 3.6.3.2.1 --- Protein extraction from rice calli and leaves --- p.124 / Chapter 3.6.3.2.2 --- Protein extraction from immature and mature rice seeds --- p.124 / Chapter 3.6.3.3 --- Tricine SDS-PAGE --- p.125 / Chapter 3.6.3.4 --- Western blot analysis --- p.125 / Chapter 3.6.4 --- Detection at metabolite level --- p.126 / Chapter 3.6.4.1 --- Isoprenoids extraction from plant materials --- p.126 / Chapter 3.6.4.2 --- UPLC analysis for isoprenoid identification --- p.127 / Chapter 3.6.5 --- Detection of promoter activity --- p.128 / Chapter 3.6.5.1 --- Histochemical staining of GUS activity --- p.128 / Chapter 3.6.5.1.1 --- Histochemical staining --- p.128 / Chapter 3.6.5.1.2 --- Plant tissue fixation for microscopic observation --- p.128 / Chapter 3.6.5.2 --- GUS activity assay --- p.129 / Chapter 3.6.5.2.1 --- Protein extraction and quantitation with Bio-Rad protein assay --- p.129 / Chapter 3.6.5.2.2 --- G US activity assay --- p.130 / Chapter CHAPTER 4. --- RESULTS --- p.131 / Chapter 4.1 --- Tissue-specificity and endosperm specific expression of rice psy1 and Psy2 --- p.131 / Chapter 4.1.1 --- Construction of gene cassettes for study in Ospsy1 and Ospsy2 --- p.133 / Chapter 4.1.2 --- Rice transformation --- p.135 / Chapter 4.1.3 --- Transgene detection --- p.137 / Chapter 4.1.3.1 --- Genomic DNA PCR screening --- p.137 / Chapter 4.1.3.2 --- Southern blot analysis --- p.139 / Chapter 4.1.3.2.1 --- Southern blot analysis on transgenic rice calli --- p.141 / Chapter 4.1.3.2.2 --- Southern blot analysis on regenerated rice --- p.143 / Chapter 4.1.4 --- Detection of transgene expression --- p.149 / Chapter 4.1.4.1 --- Northern blot analysis on immature transgenic seed --- p.149 / Chapter 4.1.4.2 --- Western blot analysis on transgenic rice tissues --- p.152 / Chapter 4.1.4.2.1 --- Antibody production --- p.152 / Chapter 4.1.4.2.2 --- Western blot analysis of transgenic rice calli --- p.155 / Chapter 4.1.4.2.3 --- Western blot analysis of transgenic rice leaves --- p.157 / Chapter 4.1.4.2.4 --- Western blot analysis of immature transgenic rice seeds --- p.160 / Chapter 4.1.5 --- Detection of OsPSYs activity at metabolite level --- p.163 / Chapter 4.1.5.1 --- UPLC analysis on transgenic rice tissues --- p.163 / Chapter 4.1.5.1.1 --- Carotenoid profiling of transgenic rice calli --- p.163 / Chapter 4.1.5.1.2 --- Carotenoid profiling of transgenic rice leaves --- p.168 / Chapter "4.1.5.1,3" --- Carotenoid profiling of mature transgenic rice seeds --- p.172 / Chapter 4.2 --- Promoter analysis of modified rice psy1 promoter --- p.176 / Chapter 4.2.1 --- Construction of gene cassettes for promoter analysis --- p.178 / Chapter 4.2.2 --- Rice transformation --- p.180 / Chapter 4.2.3 --- Transgene detection --- p.180 / Chapter 4.2.3.1 --- Genomic DNA PCR screening --- p.180 / Chapter 4.2.3.2 --- Southern blot analysis --- p.185 / Chapter 4.2.3.2.1 --- Southern blot analysis of regenerated rice --- p.186 / Chapter 4.2.4 --- Detection of promoter activity --- p.196 / Chapter 4.2.4.1 --- Promoter activity in transgenic rice leaves --- p.196 / Chapter 4.2.4.1.1 --- Histochemical staining of GUS --- p.196 / Chapter 4.2.4.1.2 --- G US activity assay --- p.199 / Chapter 4.2.4.2 --- Promoter activity in transgenic immature seeds --- p.202 / Chapter 4.2.4.2.1 --- Histochemical staining of GUS --- p.202 / Chapter 4.2.4.2.2 --- GUS activity assay --- p.206 / Chapter CHAPTER 5. --- DISCUSSIONS --- p.209 / Chapter 5.1 --- Tissue-specificity and endosperm specific expression of rice psyl and psy2 --- p.209 / Chapter 5.1.1 --- OsPSYl and OsPSY2 activities in rice calli --- p.209 / Chapter 5.1.2 --- OsPSYl and OsPSY2 activities in rice leaves --- p.210 / Chapter 5.1.3 --- OsPSYl and OsPSY2 activities in rice seeds --- p.211 / Chapter 5.2 --- Analysis of modified rice psyl promoter --- p.213 / Chapter 5.3 --- Future prospects of Golden Rice --- p.214 / Chapter CHAPTER 6. --- CONCLUSIONS --- p.217 / REFERENCES --- p.219

No description available.

Thesis (M.Sc.). / Written for the Dept. of Plant Science. Title from title page of PDF (viewed 2008/05/29). Includes bibliographical references.

Thesis (M.S.)--West Virginia University, 1999. / Title from document title page. Document formatted into pages; contains iv, 64 p. : ill. Vita. Includes abstract. Includes bibliographical references (p. 61-64).

Thesis (PhD)--Macquarie University, Division of Environmental & Life Sciences, Dept. of Biological Sciences, 2005. / Supplementary material to figures contained on DVD only available with manuscript. Bibliography: p. 221-244.

Thesis (M.S.)--Eastern Illinois University, 2009. / Includes bibliographical references (leaves 66-72).

Thesis (Ph. D.)--University of Oulu. / Includes bibliographical references.

The basic helix-loop-helix (bHLH) family oftranscription factors consists of proteins involved in cellular proliferation and differentiation. The HLH structure plays a key role in protein-protein dimerization and with the DNA target sites, referred to as E boxes containing the consensus DNA sequence CANNTG. One class of mammalian class I bHLH proteins includes products of the E2A gene, which result from alternative splicing (E12, E47, and ITF), E2-2 and HEB. E2A proteins have also been detected in most cell lines with high levels of expression in lymphoid- and pancreatic cells. It has also been demonstrated that E2A is required for B cell maturation, T cell development and has been shown to function as tumor suppressors. To date, an E2A-interacting bHLH transcription factor largely restricted to activated B lymphocytes, called ABF -1, was isolated using the two-hybrid system. ABF -1 is the only B cell restricted bHLH protein isolated. ABF-1/E2A heterodimers have been detected in B lymphocytes. In these studies, the mapping of the ABF-1 promoter and the critical 5' regulatory elements that control ABF-1 gene expression were analyzed through 5' deletional analysis. 5' -DNA flanking pieces of the promoter region were created through PCR and inserted into a promoterless cloning vector containing the firefly luciferase reporter gene. RT -PCR analysis and anchored PCR was utilized to demonstrate the transcriptional activity of the - promoter region of the ABF-1 gene. Transient transfections were completed to determine critical regulatory sequences. The promoter location was confirmed through computer analysis of the nucleotide sequence and deletional analysis.