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E . coli 에 의해 생산된 재조합 사람 프리트롬빈 2 의 정제 및 활성화
소인섭,이승철,김수원,함경수,김지영 ( In Seop So,Seung Cheol Lee,Soo Won Kim,Kyung Soo Hahm,Ji Young Kim ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.1
Human prethrombin 2 was purified from a recombinant E. coli cells in which human prethrombin 2 was produced as insoluble aggregates, amounting to 5 to 10% of the total bacterial proteins. Isolation of inclusion bodies and the subsequent solubilization of the insoluble proteins in a denaturing solution containing sodium dodecyl sulfate and dithiothreitol yielded good recovery of prethrombin 2 with at least 75% purity. Prethrombin 2 was further purified by two passages of Sephacryl S-200 gel chromatography. The purified prethrombin 2 was reacted with the polyclonal antibodies against human prothrombin. The N-terminal sequence of the prethrombin 2 was same as that of the native human prethrombin 2. The purified prethrombin 2 was renatured by stepwise removal of the denaturants by slow dialysis. The renatured prethrombin 2 was activated to thrombin by ecarin, a prothrombin activator from Echis carinatus and the thrombin activity generated was detected by a chromogenic assay. This study showed that recombinant prethrombin 2 produced in E. coli could be converted to thrombin.
국내산 프로폴리스 추출물의 기능성 플라보노이드 성분 및 함량 비교 연구
윤찬건(Yun, Chan-Gun),조세희(Cho, Sehee),김성준(Kim, Seong-Jun),이중헌(Lee, Jung-Heon),정강현(Jung, Gang-Hyeon),김지인(Kim, Jee-In),소인섭(So, In-Seop),박영현(Park, Young-Hyeon),임재환(Lim, Jae-Hwan) 한국과학영재교육학회 2021 과학영재교육 Vol.13 No.3
프로폴리스에 함유된 다양한 플라보노이드 성분은 생산 지역에 따라 함량이 다르기 때문에 그 표준화가 어렵다. 본 연구에서는 먼저 프로폴리스의 추출방법을 최적화하고, 두 번째로 프로폴리스 추출물의 지역별 플라보노이드 함량의 차이를 비교하고 주요 성분의 효능을 비교하였다. 지역별 프로폴리스의 총 폴리페놀 함량을 측정하고 DPPH assay를 통해 항산화 효능을 확인하였다. HPLC 분석을 통하여, 주요 플라보노이드 성분인 quercetin, caffeic acid, p-coumaric acid, pinocembrin, kaempferol, galangin 및 chrysin의 프로폴리스 내 함유량을 정량하였다. 마지막으로 β-hexosaminidase release assay를 통해 quercetin의 항알레르기 효능을 확인하였다. 경상북도 안동, 봉화, 군위에서 생산된 프로폴리스 추출물의 항산화 효과는 각각 80.2%, 72.4%, 54.5%였다. 프로폴리스 추출물의 플라보노이드 함량은 지역별로 차이가 있었으며 quercetin, pinocembrin, galangin, chrysin의 함량이 공통적으로 가장 높았다. Quercetin의 β-hexosaminidase 방출이 양성대조군의 약 25%로 나타나 알레르기 억제 효능을 확인하였다. 본 연구에서 프로폴리스 추출법의 표준화와 더불어 지역별 프로폴리스 추출물의 플라보노이드 함량과 각 성분별 효능을 비교하였으며, 플라보노이드에 의한 천연 항알레르기제의 활용 가능성을 제시하였다. Standardization of the various flavonoid components contained in propolis is very difficult because the contents vary depending on the production regions. In this study, first, the extraction method of propolis was optimized, secondly, the differences in flavonoids contents by region of propolis extract were compared and the biological effects of major components were compared. The total polyphenol contents of propolis from different regions were measured and the antioxidant effects were confirmed by DPPH assay. Through HPLC analysis, the contents of major flavonoids including quercetin, caffeic acid, p-coumaric acid, pinocembrin, kaempferol, galangin, and chrysin in propolis were quantified. Finally, the antiallergic effect of quercetin was confirmed through a beta-hexosaminidase release assay. The antioxidant effects of propolis extracts produced in Andong, Bonghwa, and Gunwi in Gyeongsangbuk-do were 80.2%, 72.4%, and 54.5%, respectively. The contents of flavonoids in the propolis extract varied by production region, and quercetin, pinocembrin, galangin, and chrysin were the highest contents in common. Inhibition activity of beta-hexosaminidase release by quercetin showed about 75% reduction efficacy compared to the positive control group. In this study, we compared the contents of flavonoids in propolis extracts from various regions and the effects of each component and thus we suggest the semi-standardization of the propolis extraction method and the possibility of a natural antiallergic agent by flavonoids.
TRANSFORMATION OF POPULUS TREMULOIDES USING AGROBACTERIUM RHIZOGENES.
U,Zang-Kual,Koh,Young-Hwan,So,In-Seop,Riu,Key-Zung,Lee,Sun-Joo 제주대학교 방사능이용연구소 1994 연구보고 Vol.8 No.-
Several factors affecting Arhizogenes-mediated transformation of Populus were studied. The leaf section of Populus was more sensitive to kanamycin used for selection of transformant than the stem section. The soaking period for inoculation did not affect gall formation up to 2 hours. The optimum concentration of acetosyringone and pH of bacterial culture medium for inoculation were 50μM and 5.5, respectively. One day cocultivation after inoculation gave highest transformation rate. The visible hairy roots were formed from the transformed leaf sections within 3 weeks after culture on both of the media with and without growth regulators. The plantlets were regenerated from the infected leaf sections within 6 weeks after culture on the medium containing 0.005mg/1 of NAA and 0.5mg/1 of BA. The expression of the introduced opine genes in the plantlets were confirmed by analysis of agropine and mannopine.