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
- 2017.12
- 수록면
- 265 - 270 (6page)
- DOI
- 10.7841/ksbbj.2017.32.4.265
이용수
초록· 키워드
오류제보하기
Previously, we constructed the recombinant plasmid which containing human epidermal growth factor and collagen binding domain for overproduction of fused biofunctional protein. However, this fusion protein was expressed in Escherichia coli as insoluble protein form in cytoplasm. Therefore, effective denaturation and dialysis process is critical for solubilization and refolding in protein purification process. We attempted several chemicals and buffer conditions for induction, dialysis, and solubilization. Using lactose instead of isopropyl β-D-1-thiogalactopyranoside, expression of target protein was induced. 20 mM tris-HCl buffer for dialysis was suitable for the activity and soluble form of fusion protein. Furthermore, for the solubility of expressed inclusion protein, we conducted with various pH conditions and concentrations of urea and guanidine hydrochloride. The efficient solubility of inclusion body form of fusion protein was showed at alkaline pH condition containing urea.
목차
Abstract
1. INTRODUCTION
2. MATERIALS AND METHOD
3. RESULTS AND DISCUSSION
4. CONCLUSION
REFERENCES
참고문헌 (12)
참고문헌 신청
Arora, D. and N. Khanna (1996) Method for increasing the yield of properly folded recombinant human gamma interferon from inclusion bodies. J. Biotechnol. 52: 127-133.
Baneyx, F. (1999) Recombinant protein expression in Escherichia coli. Curr. Opin. Biotechnol. 10: 411-421.
Kajava, A. V., S. N. Zolov, A. E. Kalinin, and M. A. Nesmeyanova (2000) The net charge of the first 18 residues of the mature sequence affects protein translocation across the cytoplasmic membrane of gram-negative bacteria. J. Bacteriol. 182: 2163-2169.
Kane, J. F. and D. L. Hartley (1988) Formation of recombinant protein in inclusion body in Escherichia coli. Trends Biotechnol. 6: 95-101.
Kim D. G., M. K. Min, S. C. Ahn, J. K. Kim, and I. S. Kong (2009) Expression of fusion protein containing human epidermal growth factor and the collagen-binding domain of Vibrio mimicus metalloprotease. Biotechnol. Lett. 31: 259-264.
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UCI(KEPA) : I410-ECN-0101-2018-470-001673597