Title page for etd-0205110-152503


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URN etd-0205110-152503
Author Shih-hsien Chao
Author's Email Address No Public.
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Department Biological Sciences
Year 2009
Semester 1
Degree Master
Type of Document
Language zh-TW.Big5 Chinese
Title Cloning and Expression of Arabidopsis endo-1,4-β-glucanase in Pichia pastoris
Date of Defense 2010-01-13
Page Count 69
Keyword
  • Pichia pastoris
  • endo-1, 4-β-glucanase
  • Abstract In recent years. as industrialized society developed. people have made a lot of environment pollution problems owing to overusing fossil fuel, moreover. fossil fuel is going to deplete. As the result. the study of the substitute energy is promoted.
    Ligonocellulose (lignin, cellulose and hemicelluloses are included) are the most plentiful renewable resources in the nature, and it have high economic values which extensively use on food, paper and energy. Recent years, it is a hot issue that decomposing cellulose into the minimum unit called glucose, and further, fermenting into alcohol to generate biomass energy.
    This research goal is cloning Arabidopsis endo-1,4-β-glucanase and transfer to Escherichia coli(DH5α). Selection pPICZαA the success transferr DNA Fragment. Then to sequence and because of cell in pPICZαA have expression in Pichia. pastoris. AOX1 promoter have Mass productions Arabidopsis endo-1,4-β-glucanase gene in Pichia pastoris. Carries on the extracellular expression by the methyl alcohol induction way. Can obtain recombinant DNA protein. However the Western blotting analysis demonstration has this enzyme active protein At4g11050 pellet Molecular weight about 89 KDa. Again by way of congo red and Dye-CMC assay activeness of the examination enzyme protein. The result discovers At4g11050 in the pellet cell activity compares with negative control has the obvious activity.
    Advisory Committee
  • Ching-Mei Hsu - chair
  • Mang-Jye Ger  - co-chair
  • Zin-Huang Liu - advisor
  • Files
  • etd-0205110-152503.pdf
  • indicate not accessible
    Date of Submission 2010-02-05

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