Heat shock proteins of plants were main protein in some evolution stress.Expression of some heat shock proteins had a certain relationship with plant tolerance.The nucleotide sequence of a cDNA cloned from Arabidopsis thaliana ecotype RLD was determined,and the corresponding amino sequence deduced.The open reading frame encodes a DnaJ protein of 442 residues with a molecular mass of 47 ku.The predicted sequence contains regions homologous to the J-and cysteine-rich domains of Escherichia coli DnaJ.The cDNA fragment directionally cloned into the vector pET32α,over-expression of DnaJ in E.coli,the E.coli could grow on LB containing 0.5 mol/L NaCl.

Over-expression of Arabidopsis DnaJ(Hsp40) Contributes to NaCl-stress Tolerance of Escherichia coli

...................................................................................................................................................................... i Acknowledgements ............................................................................................................................................. iii Abbreviations ......................................................................................................................................................... iv List of Figures......................................................................................................................................................... vi List of Tables ......................................................................................................................................................... vii Chapter 1: Introduction ...................................................................................................................................... 1 1.1 General Introduction to CPR5 ............................................................................................................ 1 1.2 Phytohormones Signalling ................................................................................................................... 3 1.3 Plant Germination and Development .............................................................................................. 5 1.4 Plant Senescence ...................................................................................................................................... 7 1.5 Resistance and Hypersensitive Response-Mediated PCD ....................................................... 8 1.6 Reactive Oxygen Species (ROS) .......................................................................................................11 1.7 CPR5 Molecular Function: Protein Localization and Structural Analysis .......................12 1.7 CPR5 and Cell Cycle Involvement ....................................................................................................14 1.8 Concluding Remarks.............................................................................................................................15 Chapter 2: Materials and Methods ...............................................................................................................17 Chemicals used ...............................................................................................................................................17 2.1 General Use Protocols: .........................................................................................................................17 2.1.1 Bacterial Propagation ................................................................................................................17 2.1.2 Preparation of Plasmid DNA (Alkaline Lysis Miniprep) ..............................................18 2.1.3 Agarose Gel Electrophoresis ...................................................................................................18 2.1.4 Preparation of Chemically Competent Bacterial Strains .............................................19 2.2 Cloning 19 2.2.1 PCR Amplification of cDNA ......................................................................................................19 2.2.2 Restriction Digestion and DNA Ligation.............................................................................20 2.2.3 Bacterial Chemical Transformation .....................................................................................20 2.2.4 Bacterial Colony PCR ..................................................................................................................21 2.2.5 DNA Sequencing ...........................................................................................................................21 2.3 SDS PAGE and Western Blot Analysis ...........................................................................................22 2.3.1 Protein Extraction and Sodium Dodecyl Sulfate (SDS) Polyacrylamide Gel Electrophoresis (PAGE) ...........................................................................................................22 2.3.2 Transfer of Protein onto PVDF Membrane........................................................................24 2.4 Yeast-Two-Hybrid (Y2H) ....................................................................................................................24 2.4.1 Generating Bait Plasmids for Y2H “Mate and Plate” Library Screening ................25

Characterization of Arabidopsis thaliana CPR5 via the elucidation of interacting protein partners : a thesis presented in partial fulfilment of the requirements for the degree of Masters of Science in Biochemistry at Massey University, Palmerston North, New Zealand

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Characterization of Arabidopsis thaliana CPR5 via the elucidation of interacting protein partners : a thesis presented in partial fulfilment of the requirements for the degree of Masters of Science in Biochemistry at Massey University, Palmerston North, New Zealand