| URN |
etd-0715113-214714 |
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| Author |
Meng-Yao Lee |
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| Author's Email Address |
No Public. |
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| Statistics |
This thesis had been viewed 5573 times. Download 660 times. |
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| Department |
Biological Sciences |
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| Year |
2013 |
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| Semester |
1 |
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| Degree |
Master |
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| Type of Document |
|
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| Language |
zh-TW.Big5 Chinese |
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| Title |
Expression of Fractalkine chemokine domain by baculovirus expression system |
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| Date of Defense |
2013-07-11 |
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| Page Count |
66 |
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| Keyword |
Fractalkine
His tag
baculovirus
|
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| 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. |
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| Advisory Committee |
Song-Tay Lee - chair
Jin-Ching Lee - co-chair
Jong-Kang Liu - advisor
Hung Wu - advisor
|
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| Files |
Indicate in-campus at 1 year and off-campus access at 5 year. |
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| Date of Submission |
2013-08-15 |
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