DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | Lee, Young-Hoon | - |
dc.contributor.advisor | 이영훈 | - |
dc.contributor.author | Kim, Dae-Hee | - |
dc.contributor.author | 김대희 | - |
dc.date.accessioned | 2011-12-13T05:00:44Z | - |
dc.date.available | 2011-12-13T05:00:44Z | - |
dc.date.issued | 1996 | - |
dc.identifier.uri | http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=105580&flag=dissertation | - |
dc.identifier.uri | http://hdl.handle.net/10203/32724 | - |
dc.description | 학위논문(석사) - 한국과학기술원 : 화학과, 1996.2, [ iv, 56 p. ] | - |
dc.description.abstract | Tenecin 3 is one of the antifungal proteins present in hemolymph of larvae of Tenebrio molitor constitutively. Since large quantity of natural tenecin 3 was not available for study of antifungal action, an expression system for recombinant tenecin 3 in Escherichia coli was developed. The T7 expression system was used for overproduction of recombinant tenecin 3 protein. For expression of intact tenecin 3 and precursor tenecin 3, the DNA fragments of cDNA amplified by PCR were subcloned into pET-3a vector and pET-21a(+) vector. However, the corresponding recombinant protein expressed from the resulting plasmid in E. coli was not detected by protein SDS-PAGE. To overcome problems originated from low expression of tenecin 3, expression of the recombinant proteins as fusion to His tag and/or as secretory proteins to periplasm was attempted. From the plasmid expressing His tag tenecin 3 fusion protein, the recombinant protein was overproduced and purified by $Ni^{2+}$-chelating affinity chromatography. His tag tenecin 3 fusion protein showed good repression on the growth of the fungus Candida albicans like natural tenecin 3 did. In case of the plasmid for secretion of tenecin 3 to the periplasm, the recombinant tenecin 3 protein showed no repression on the growth of C. albicans although it was successfully overproduced and secreted. Using His tag tenecin 3 fusion expression system, the effect of the deletion of the tenecin 3 sequence on antifungal action was examined to obtain information about amino acids essential for antifungal activity. Several plasmids expressing serial N-terminal truncated proteins and C-terminal truncated proteins, respectively, were constructed, from which the truncated tenecin 3 proteins were purified as fusion to His tag and tested for antifungal activity against C. albicans. The results showed that tenecin 3 has, at least, two active domains; one of them locates in the C-terminal half region and another locates in the N-terminal half region. | eng |
dc.language | eng | - |
dc.publisher | 한국과학기술원 | - |
dc.subject | Escherichia coli | - |
dc.subject | Expression | - |
dc.subject | Antifungal protein | - |
dc.subject | Insect | - |
dc.subject | Active site | - |
dc.subject | 활성부위 | - |
dc.subject | 대장균 | - |
dc.subject | 발현 | - |
dc.subject | 항진균 단백질 | - |
dc.subject | 곤충 | - |
dc.title | Expression of an insect antifungal protein in Escherichia coli | - |
dc.title.alternative | 곤충 유래 항진균 단백질의 대장균내 발현 | - |
dc.type | Thesis(Master) | - |
dc.identifier.CNRN | 105580/325007 | - |
dc.description.department | 한국과학기술원 : 화학과, | - |
dc.identifier.uid | 000943051 | - |
dc.contributor.localauthor | Lee, Young-Hoon | - |
dc.contributor.localauthor | 이영훈 | - |
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