지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2011.6
- 수록면
- 199 - 205 (7page)
이용수
초록· 키워드
오류제보하기
Using tetrameric β-galactosidase as a model protein, anchoring motives were screened in Bacillus subtilis spore display system. Eleven spore coat proteins were selected considering their expression levels and the location in the spore coat layer. After chromosomal single-copy homologous integration in the amyE site of Bacillus subtilis chromosome, cotE and cotG were chosen as possible spore surface anchoring motives with their higher whole cell β-galactosidase activity. PAGE and Wester blot of extracted fraction of outer layer of purified spore, which express CotE-LacZ or CotG-LacZ fusion verified the existence of exact size of fusion protein and its location in outer coat layer of purified spore. β-galactosidase activity of spore with CotE-LacZ or CotG-LacZ fusion reached its highest value around 16~20 h of culture time in terms of whole cell and purified spore. After intensive spore purification with lysozyme treatment and renografin treatment, spore of BJH135, which expresses CotE-LacZ, retained only 1~2% of its whole cell β-galactosidase activity. Whereas spore of BJH136, which has cotG-lacZ cassette in the chromosome, retained 10~15% of its whole cell β-galactosidase activity, proving minor perturbation of CotG-LacZ, when incorporated in the spore coat layer of Bacillus subtilis compared to CotE-LacZ. Usage of Bacillus subtilis WB700, of which 7 proteases are knocked-out and thereby resulting in 99.7 % decrease in protease ctivity of the host, did not prevent the proteolytic degradation of spore surface expressed CotG-LacZ fusion protein.
목차
Abstract
1. 서론
2. 재료 및 방법
3. 결과 및 고찰
4. 결론
References
참고문헌 (22)
참고문헌 신청
Charbit, A / 1988 / Versatility of a vector for expressing foreign polypeptides at the surface of gram-negative bacteria / Gene 70:181~189
Boder, E. T / 1997 / Yeast surface display for screening combinatorial polypeptide libraries / Nat. Biotechnol 15:553~557
Jung, H. C / 1998 / Surface display of Zymomonas mobilis levansucrase by using the ice-nucleation protein of Pseudomonas syringae / Nat. Biotechnol 16:576~580
Georgiou, G / 1997 / Display of heterologous proteins on the surface of microorganisms: from the screening of combinatorial libraries to live recombinant vaccines / Nat. Biotechnol 15:29~34
Richins, R. D / 1997 / Biodegradation of organophosphorus pesticides by surfaceexpressed organophosphorus hydrolase / Nat. Biotechnol 15:984~987
Boder, E. T / 1997 / Yeast surface display for screening combinatorial polypeptide libraries / Nat. Biotechnol 15:553~557
Jung, H. C / 1998 / Surface display of Zymomonas mobilis levansucrase by using the ice-nucleation protein of Pseudomonas syringae / Nat. Biotechnol 16:576~580
Georgiou, G / 1997 / Display of heterologous proteins on the surface of microorganisms: from the screening of combinatorial libraries to live recombinant vaccines / Nat. Biotechnol 15:29~34
Richins, R. D / 1997 / Biodegradation of organophosphorus pesticides by surfaceexpressed organophosphorus hydrolase / Nat. Biotechnol 15:984~987
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UCI(KEPA) : I410-ECN-0101-2014-400-002390557