Title page for etd-0715113-214714


[Back to Results | New Search]

URN etd-0715113-214714
Author Meng-Yao Lee
Author's Email Address No Public.
Statistics This thesis had been viewed 5385 times. Download 398 times.
Department Biological Sciences
Year 2013
Semester 1
Degree Master
Type of Document
Language zh-TW.Big5 Chinese
Title Expression of Fractalkine chemokine domain by baculovirus expression system
Date of Defense 2013-07-11
Page Count 66
Keyword
  • Fractalkine
  • His tag
  • baculovirus
  • Abstract Baculovirus expression system has often been utilized to express recombinant proteins.The advantages of this system include high efficiency, ability to express proteins in manytypes of cells, and production of proteins with correct conformation and posttranslational modifications. Fractalkine is a membrane protein mainly exerting its effects through binding of extracellular chemokine domain to the CX3CR1 receptor. It regulates proliferation, differentiation, and apoptosis of cells. It also plays an important role in cardiovascular diseases. The goal of this thesis is to construct a baculovirus vector system to express fractalkine chemokine domain for further functional studies. The vector system is engineered to contain a secretion signal peptide sequence and two affinity tags, polyhistidine tag for purification and myc tag for immunoblotting. Using this system, it was found that fractalkine chemokine domain is able to express in HEK293 and HeLa cells, whereas THP-1 monocyte and Jurkat T cells are without effect. The expressed recombinant protein could be detected at 60 h after viral infection and the majority appeared to be produced after 72 h. In addition, the fractalkine chemokine domain produced in HEK293 cells was able to purified from the medium through polyhistidine tag pulldown. These results indicate that a highly efficient protein expression system with advantages of easy detection and purification has been established and this system is a valuable tool in structural and functional studies of proteins.
    Advisory Committee
  • Song-Tay Lee - chair
  • Jin-Ching Lee - co-chair
  • Jong-Kang Liu - advisor
  • Hung Wu - advisor
  • Files
  • etd-0715113-214714.pdf
  • Indicate in-campus at 1 year and off-campus access at 5 year.
    Date of Submission 2013-08-15

    [Back to Results | New Search]


    Browse | Search All Available ETDs

    If you have more questions or technical problems, please contact eThesys