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대장균에서 재조합 인간 Interleukin 4 의 생산 , 정제 및 면역조절활성의 측정
양영,윤석란,이충은,변광호 ( Young Yang,Suk Ran Yoon,Choong Eun Lee,Kwang Ho Pyun ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.1
The recombinant human interleukin 4 (rhIL-4) has been over expressed in E. coli transformed with expression vector pET-3b containing bacteriophage T7 promoter, into which the hIL-4 cDNA was subclond. The insolubility of the recombinant protein offered an advantage of purification in only a few steps. The recombinant human IL-4 was refolded using reduced/oxidized glutathione to restore the proper conformation and purified to homogeneity by one passage over ion exchange column. The purified protein was shown as a single band on SDS-PAGE. The refolded rhIL-4 was characterized by nucleotide sequence analysis and bioassays. The purified rhIL-4 has biological activities on B cell proliferation and induction of B cell differentiation antigen, CD23, which strongly indicates that the protein is folded correctly.
대장균에서 재조합 인간 Interleukin 6의 대량생산 및 그 생물학적 활성의 측정
양영,강형식,나우진,이충은,변광호 ( Young Yang,Hyung Sik Kang,Woo Jin Na,Choong Eun Lee,Kwang Ho Pyun ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.2
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.
가열조리가 복어 추출물 함질소화합물의 조성에 미치는 영향
양영,한영실,변재형,Yang, Yeong,Han, Yeong-Sil,Pyeun, Jae-Hyeung 한국식품조리과학회 1990 한국식품조리과학회지 Vol.6 No.2
Globefish, Fugu xanthopterus, known to have a severe toxin, is one of the favorite food in Korea and Japan when the toxic part is removed. In this paper, the effect of cooking on the composition of nitrogenous components in the extractives from globefish cooked investigated and the changes of the taste compounds originated from the nitrogenous components in the extractives were discussed. When the sample fish was analysed for general composition, drip amount and pH by the different methods of thawing, the method effective method was the running water thawing. Total nitrogen content in raw globefish and the frozen globefish was not different, and the nitrogen content was increased with the heat treatment. It seemed that the nitrogen content was higher in the extract from the boiled globefish than that of the steamed globefish. Taurine, lysine, glycine and alanine were occupied about 70% of the total free amino acids. Total free amino acid content was higher in the extracts from the frozen sample than those from the raw sample. The amount of free amino acids was increased when the globefish soup cooked under the direct-heat cooking than in the microwave oven-heat cooking. Among nucleotides in the extracts from the thawed and cooked fishes, IMP and inosine contents were increased, and the both components were decreased with the heating time and by the heating method. Tne content of total creatinine-nitrogen were 50% of the total nitrogen content of the extracts, but the concentration of glycinebetaine, TMA and TMAO were only a few amounts. It could be concluded that total creatinine components, including free amino acids such as taurine, lysine, glycine and alanine, and IMP might be the important components contributing to the taste of the cooked globefish.
NF-κB전사인자가 coactivator을 통하여 VDUP1 발현을 억제하는 기전
양영,김근영 대한암예방학회 2007 Journal of cancer prevention Vol.12 No.1
Vitamin D3 up-regulated protein 1 (VDUP1) is a stress responsive gene that is up-regulated by 1,25 (OH)2D3, an active form of vitamin D3. It has been reported that the expression of VDUP1 is reduced in many tumor cells and that the enforced expression of VDUP1 inhibits the proliferation of tumor cells. We therefore consider VDUP1 as a tumor suppressor. In the present study, we found that VDUP1 is down- regulated by NF-κB transcription factor in human gastric cancer cell line AGS. On the contrary to VDUP1, NF-κB enhances the growth and proliferation of tumors and its expression is up-regulated in various tumors. To determine the regulatory mechanism by which NF-κB suppresses the transcription of VDUP1, the various deletion constructs of VDUP promoter were designed. As a result of promoter analysis, we found that NF-κB did not bind to VDUP1 promoter region, but it could inhibit the recruitment of coactivators (PCAF, p300 and CBP) to NF-Y, which plays a critical role in the increase of VDUP1 transcription. SAHA-mediated the recruitment of coactivators to NF-Y was also inhibited by NF-κB. These results imply that activation of NF-κB suppresses VDUP1 gene induction by SAHA through preventing the recruitment of coactivators to NF-Y in human gastric cancer cells. (Cancer Prev Res 12, 14-20, 2007) Key Words: VDUP1, NF-κB, p300, pCAF