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Cloning and analysis of promoter regulating the expression of a purple acid phosphatase.January 2001 (has links)
Zhang Siyi. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 97-109). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.iii / List of Tables --- p.vii / List of Figures --- p.viii / List of Abbreviations --- p.x / Chapter Chapter 1: --- General Introduction --- p.1 / Chapter Chapter 2: --- Literature Review --- p.3 / Chapter 2.1 --- Phosphorus and higher plants --- p.3 / Chapter 2.1.1 --- Phosphorus is a macronutrient in higher plants --- p.3 / Chapter 2.1.2 --- The forms of phosphorus in plant cells --- p.3 / Chapter 2.1.3 --- Phosphorus compartments and pools in plant cells --- p.6 / Chapter 2.2 --- The acquisition of phosphorus in higher plants --- p.8 / Chapter 2.2.1 --- The forms of phosphorus absorbed by higher plants --- p.8 / Chapter 2.2.2 --- Soil phosphorus bioavailability --- p.9 / Chapter 2.2.3 --- Uptake and transportation of phosphorus --- p.10 / Chapter 2.3 --- Adaptive responses of higher plants to phosphorus deficiency --- p.11 / Chapter 2.3.1 --- Phosphorus homeostasis --- p.12 / Chapter 2.3.2 --- Enhancement of phosphorus uptake --- p.14 / Chapter 2.3.3 --- Phosphorus scavenging and recycling --- p.16 / Chapter 2.4 --- Regulation of gene expression under phosphorus starvation --- p.18 / Chapter 2.5 --- Acid phosphatase and purple acid phosphatase in plants --- p.22 / Chapter 2.5.1 --- Acid phosphatases --- p.22 / Chapter 2.5.2 --- Purple acid phosphatase (PAP) --- p.26 / Chapter Chapter 3: --- Hypothesis --- p.31 / Chapter Chapter 4: --- Materials and Methods --- p.33 / Chapter 4.1 --- Materials --- p.33 / Chapter 4.1.1 --- Chemicals --- p.33 / Chapter 4.1.2 --- Plant materials --- p.33 / Chapter 4.1.3 --- Plasmid vectors and bacterial strains --- p.33 / Chapter 4.1.4 --- DNA sequencing --- p.34 / Chapter 4.1.5 --- Softwares: --- p.34 / Chapter 4.2 --- Methods: --- p.35 / Chapter 4.2.1 --- Survey of PAP occurrence in higher plants --- p.35 / Chapter 4.2.2 --- Determination of multi-gene family and gene copy number of PAPin tomato genome --- p.40 / Chapter 4.2.3 --- Effect of environmental Pi on the morphology of tomato and APase induction --- p.43 / Chapter 4.2.4 --- PAP expression in tomato seedlings growing at different Pi concentrations --- p.46 / Chapter 4.2.5 --- Genomic library construction and PAP promoter isolation --- p.48 / Chapter 4.2.6 --- PAP promoter activity assay by transient expression of reporter gene..… --- p.52 / Chapter Chapter 5: --- Results --- p.56 / Chapter 5.1 --- Identification of PAP gene in higher plants --- p.56 / Chapter 5.1.1 --- Design of primers and total RNA extraction --- p.56 / Chapter 5.1.2 --- RT-PCR --- p.57 / Chapter 5.1.3 --- Further investigation of PAP homologous sequences in monocotyledons --- p.60 / Chapter 5.2 --- Determination of multi-gene family and gene copy number of tomato PAP gene (TPAP 1) --- p.62 / Chapter 5.2.1 --- Determination of TPAP 1 copy number --- p.62 / Chapter 5.2.2 --- Determination of tomato PAP multi-gene family --- p.63 / Chapter 5.3 --- Effect of environmental phosphorus on the morphology of tomato seedling and APase induction --- p.66 / Chapter 5.3.1 --- Morphological changes of tomato seedlings under phosphorus starvation --- p.66 / Chapter 5.3.2 --- Acid phosphatase assays --- p.72 / Chapter 5.4 --- The phosphorus-regulated expression of tomato PAP --- Northern blot analysis --- p.74 / Chapter 5.5 --- Genomic library construction and PAP promoter isolation --- p.76 / Chapter 5.6 --- PAP promoter sequence --- p.79 / Chapter 5.7 --- Promoter activity assay through transient expression of reporter gene --- p.84 / Chapter 5.7.1 --- Effect of untranslation region of PAP gene --- p.84 / Chapter 5.7.2 --- Assay of PAP promoter activities regulated by phosphorus --- p.85 / Chapter Chapter 6: --- Discussion --- p.88 / Chapter 6.1 --- The wide occurrence and high conservation of plant PAP --- p.88 / Chapter 6.2 --- Tomato as a model plant and the organization of PAP gene in genome --- p.89 / Chapter 6.3 --- Morphological changes of tomato under phosphorus starvation and the induction of APase --- p.90 / Chapter 6.4 --- Regulation of PAP in tomato --- p.92 / Chapter 6.5 --- Isolation of PAP promoter --- p.92 / Chapter 6.6 --- Assay of PAP promoter activity --- p.93 / Chapter 6.7 --- Future perspectives --- p.94 / Chapter Chapter 7: --- Conclusion --- p.95 / References --- p.97
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