Overproduction and Biological Activities of Recombinant Human Interleukin 6 Expressed in E. coli대장균에서 재조합 인간 Interleukin 6의 대량생산 및 그 생물학적 활성의 측정
- Other Titles
- 대장균에서 재조합 인간 Interleukin 6의 대량생산 및 그 생물학적 활성의 측정
- Authors
- 양영; 강형식; 나우진; 이충은; 변광호
- Issue Date
- Jan-1992
- Publisher
- 생화학분자생물학회
- Citation
- 한국생화학회지 (BMB Reports), v.25, no.2, pp 165 - 170
- Pages
- 6
- Journal Title
- 한국생화학회지 (BMB Reports)
- Volume
- 25
- Number
- 2
- Start Page
- 165
- End Page
- 170
- URI
- https://scholarworks.sookmyung.ac.kr/handle/2020.sw.sookmyung/151142
- ISSN
- 1976-6696
1976-670X
- Abstract
- Human interleukin 6 (hIL-6) is a multifunctional cytokine involved in acute phase and immune response. While a critical role of IL-6 in various immune disorders has been suggested, studies on its detailed functional mechanism are often hampered due to the low natural abundance of this cytokine. Thus, for a large scale production of hIL-6, we have made an attempt to directly express hIL-6 using pET-8c expression plasmid under the control of T7 promoter in Escherichia coli. A cDNA coding for hIL-6 without the signal sequence was amplified using PCR reaction, fused to pET-8c, and transformed into E. coli (λDE3). Upon induction with isopropyl thiogalactoside, recombinant human interleukin 6 (rhIL-6) was overexpressed as a form of inculsion bodies, with its expression level reaching over 30% of total E. coli proteins. The rhIL-6 was isolated from inclusion bodies by solubilization in 6 M guanidine hydrochloride followed by dialysis against 50 mM Tris buffer. A single passage over DEAE-Sepharose faciliated the purification of rhIL-6. The purified rhIL-6 was characterized by nucleotide sequence analysis and bioassays. The biological activity was confirmed by proliferation of B9 cells and immunoglobulin secretion of SAC-blast B cells.
- Files in This Item
-
Go to Link
- Appears in
Collections - 이과대학 > 생명시스템학부 > 1. Journal Articles
Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.