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Title page for etd-0923105-164739


URN etd-0923105-164739 Statistics This thesis had been viewed 1874 times. Download 15 times.
Author Cheng-Han Hsieh
Author's Email Address hsieh.hank@msa.hinet.net
Department Bioengineering
Year 2004 Semester 2
Degree Master Type of Document Master's Thesis
Language English Page Count 87
Title The effect of various media and inducer concentration on the expression of recombinant cystatin
Keyword
  • specific activity
  • protease
  • cystatin
  • CeCPI
  • CeCPI
  • cystatin
  • protease
  • specific activity
  • Abstract This research is base on Escherichia coli XL1-blue/pGEX-CeCPI in flask experiment to generate Cystatin。What principles should we follow to add Isopropyl-β-D-thiogalactopyranoside (IPTG) When Cystatin generates. Due to there is no assay reveals the specific activity of Cystatin. Moreover, during this experiment, we could define Cystatin as Cystatin Unit. Not only by adding Cystatin reactive volume(g)can be described. In this experiment, we learn that while we add 0.2 m IPTG to induce. No matter we use Luria Broth(LB)or Expansion Broth / Overexpression(EB/OB) Broth as the medium, the capacity of Cystatin can both reach the highest within 12 hours. But if we use EB/OB as the medium to generate Cystatin, the specific activity is 1.33 times to the one to use LB as a medium.。
    Given that induced by 0.2 mM IPTG and stored by glycerol as a reagent of freeze resistant , the specific activity will reach its highest after induced by LB or EB/OB as a medium along with glycerol excluded. Cystatin specific activity produced by EB/OB as a medium is 3.58 times than that by LB as a medium. Should glycerol is included, the specific activity will reach its highest at 3 hours after induced by LB or EB/OB as a medium. The cystatin specific activity produced by LB as a medium is 1.58 times than that without glycerol, while by EB/OB as a medium is 1.47 times that without glycerol.
    Advisor Committee
  • C. Will Chen - advisor
  • Dey-Chyi Sheu - co-chair
  • Kai-Wun Yeh - co-chair
  • Files indicate in-campus access at one year and off-campus not accessible
    Date of Defense 2005-07-27 Date of Submission 2005-09-23


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