DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | Byun, Si-Myung | - |
dc.contributor.advisor | 변시명 | - |
dc.contributor.author | Park, Seung-Kook | - |
dc.contributor.author | 박승국 | - |
dc.date.accessioned | 2011-12-12T07:34:37Z | - |
dc.date.available | 2011-12-12T07:34:37Z | - |
dc.date.issued | 1991 | - |
dc.identifier.uri | http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=61669&flag=dissertation | - |
dc.identifier.uri | http://hdl.handle.net/10203/27307 | - |
dc.description | 학위논문(박사) - 한국과학기술원 : 생물공학과, 1991.2, [ xi, 160 p. ] | - |
dc.description.abstract | The expression vector for streptokinase has been constructed from our previously cloned streptokinase coding gene($\underline{\mbox{skc}}$). Because of its deleterious effect on the host cell growth, the leader sequence of skc was removed and the leader sequence-deleted $\underline{\mbox{skc}}$ was subcloned into the vector pkk223-3, which contains the regulatable $\underline{\mbox{tac}}$ promoter and $\underline{\mbox{rrnB}}$ $T_1T_2$ transcription terminator, with a short synthetic oligonucleotide adapter. When this vector, pKS601, was expressed in $\underline{\mbox{Escherichia}}$ $\underline{\mbox{coli}}$, a 47.4 kD protein was found to be newly accumulated to about 12% of the total cellular proteins and it was identified as the streptokinase by immunoblotting with rabbit anti-streptokinase polyclonal serum. The expressed leader sequence-deleted streptokinase caused no detrimental effect to host cell physiology, which means that the deleterious effect of the wild-type $\underline{\mbox{skc}}$ had caused by its putative leader sequence, and showed no degraded 44 kD streptokinase, which is characteristic feature of streptokinase expressed from wild-type $\underline{\mbox{skc}}$. At 37℃, one-fourth of streptokinase was found in sonication-insoluble pellet, which regarded as inclusion bodies, but at 25℃, nearly all of streptokinase was found in sonication-soluble fraction. But the total activity of expressed streptokinase showed the highest values when cells had cultured and induced at 37℃, The expressed streptokinase was purified to near homogeneity using DEAE-cellulose and Sephadex G-150 column. Its specific activity was 1.3×$10^5$ CLN units/mg protein. And a series of $\underline{\mbox{E.}}$ $\underline{\mbox{coli}}$-$\underline{\mbox{Bacillus}}$ $\underline{\mbox{subtilis}}$ shuttle vectors was constructed by fusing and manipulating the $\underline{\mbox{Staphylococcus}}$ $\underline{\mbox{aureus}}$ plasmid pUB110 and $\underline{\mbox{E.}}$ $\underline{\m... | eng |
dc.language | eng | - |
dc.publisher | 한국과학기술원 | - |
dc.subject | 단백질 분리 | - |
dc.title | High-level expression of streptokinase gene in escherichia coli and development of aprE-lacUV5 hybrid promoters | - |
dc.title.alternative | 대장균에서 스트렙토키나제 유전자의 대량발현 및 aprE-lacUV5 융합 전사촉진제의 개발 | - |
dc.type | Thesis(Ph.D) | - |
dc.identifier.CNRN | 61669/325007 | - |
dc.description.department | 한국과학기술원 : 생물공학과, | - |
dc.identifier.uid | 000855138 | - |
dc.contributor.localauthor | Byun, Si-Myung | - |
dc.contributor.localauthor | 변시명 | - |
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