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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Cloning and identification of salt inducible genes in arabidopsis thaliana.

January 2000 (has links)
Chan Yee-kwan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 108-131). / Abstracts in English and Chinese. / Thesis Committee --- p.i / Abstract --- p.ii / Acknowledgments --- p.v / General Abbreviations --- p.vii / Abbreviation for Chemicals --- p.x / Table of Contents --- p.xi / List of Figures --- p.xiv / List of Tables --- p.xv / Chapter 1. --- Literature Review / Chapter 1.1 --- Salinity as a global problem --- p.1 / Chapter 1.2 --- Salinity and agriculture --- p.2 / Chapter 1.3 --- Plant adaptation to salinity --- p.4 / Chapter 1.3.1 --- Salt secretion --- p.6 / Chapter 1.3.2 --- Ion transport --- p.8 / Chapter 1.3.2.1 --- Role of H+-ATPase in salt tolerance --- p.8 / Chapter 1.3.2.2 --- Potassium and sodium uptake --- p.13 / Chapter 1.3.2.3 --- Sodium efflux --- p.15 / Chapter 1.3.3 --- Osmotic adjustment --- p.20 / Chapter 1.3.3.1 --- Accumulation of mannitol --- p.21 / Chapter 1.3.3.2 --- Accumulation of proline --- p.23 / Chapter 1.3.3.3 --- Accumulation of glycinebetaine --- p.23 / Chapter 2. --- Materials and Methods / Chapter 2.1 --- Plant materials and growth conditions --- p.26 / Chapter 2.1.1 --- Surface sterilization of Arabidopsis seeds --- p.26 / Chapter 2.1.2 --- Determination of sub-lethal inhibitory doses of sodium --- p.27 / Chapter 2.1.3 --- Growth conditions of Arabidopsis seeds for total RNA extraction --- p.27 / Chapter 2.1.4 --- NaCl dosage tests --- p.28 / Chapter 2.1.5 --- Expression kinetic tests --- p.28 / Chapter 2.2 --- Isolation of total RNAs --- p.28 / Chapter 2.3 --- Isolation of genes differentially expressed in NaCl concentration by RAP-PCR --- p.30 / Chapter 2.3.1 --- RNA fingerprinting by RAP-PCR --- p.30 / Chapter 2.3.2 --- PCR reamplificatin of RAP products --- p.31 / Chapter 2.3.3 --- Cloning of differentially expressed genes --- p.33 / Chapter 2.3.3.1 --- Ligation of inserts into pCR-Script vector and transformation --- p.33 / Chapter 2.3.3.2 --- Ligation of inserts into pBluescript II KS (+) T-vector and transformation --- p.36 / Chapter 2.3.3.3 --- Screening of recombinant plasmids --- p.37 / Chapter 2.4 --- Sequencing of differentially expressed genes --- p.39 / Chapter 2.4.1 --- DNA cycle sequencing --- p.39 / Chapter 2.5 --- Northern blot hybridization of NaCl inducible genes --- p.40 / Chapter 2.5.1 --- RNA fractionation by formaldehyde gel electrophoresis --- p.40 / Chapter 2.5.2 --- Northern blotting --- p.41 / Chapter 2.5.3 --- Preparation of single-stranded DIG-labeled PCR probes --- p.41 / Chapter 2.5.3.1 --- Isolation of Total RNA --- p.41 / Chapter 2.5.3.2 --- Primer design --- p.42 / Chapter 2.5.3.3 --- PCR amplification of single-stranded DIG PCR probes --- p.43 / Chapter 2.5.4 --- Hybridization --- p.45 / Chapter 2.5.5 --- Stringency washes --- p.46 / Chapter 2.5.6 --- Chemiluminescent detection --- p.46 / Chapter 3. --- Results / Chapter 3.1 --- Determination of sub-lethal inhibitory doses of sodium --- p.48 / Chapter 3.2 --- Isolation of total RNA from A. thaliana treated with sodium chloride --- p.48 / Chapter 3.3 --- Isolation of genes differentially expressed in sodium concentration by RNA arbitrarily primed polymerase chain reaction RAP-PCR --- p.52 / Chapter 3.3.1 --- Differential cDNA fragments identified by RAP-PCR --- p.52 / Chapter 3.3.2 --- PCR reamplification of RAP products --- p.52 / Chapter 3.3.3 --- Cloning of selected RAP-fragments --- p.62 / Chapter 3.4 --- Nucleotide sequence analysis of selected RAP PCR clones --- p.65 / Chapter 3.5 --- Expression pattern analysis of salt inducible genes by northern blot hybridization --- p.75 / Chapter 3.5.1 --- Preparation of single-stranded digoxigenin (DIG)-labeled probes --- p.75 / Chapter 3.5.2 --- Dosage response of NaCl inducible genes --- p.79 / Chapter 3.5.3 --- Expression kinetics of NaCl inducible genes --- p.80 / Chapter 4. --- Discussion / Chapter 4.1 --- Isolation of RAP-PCR targets --- p.93 / Chapter 4.2 --- Expression of NaCl inducible P450 genes --- p.94 / Chapter 4.2.1 --- Cytochrome P450 CYP73A5 --- p.97 / Chapter 4.2.2 --- Cytochrome P450 CYP83A1 --- p.98 / Chapter 4.3 --- NaCl induction gene related to post-transcriptional activities --- p.99 / Chapter 4.3.1 --- Glycine-rich RNA binding protein (BAC F3F19) --- p.100 / Chapter 4.3.2 --- Chloroplast signal recognition particle (54CP) --- p.103 / Chapter 4.4 --- Conclusion --- p.106 / References --- p.108

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