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쥐 간 미토콘드리아 막의 알데하이드 탈수소 효소의 성질 및 기능적 재조합을 위한 조건 조사
이희봉,윤자영,주충노 ( Hee Bong Lee,Ja Young Yun,Chung No Joo ) 생화학분자생물학회 1993 BMB Reports Vol.26 No.8
To know the existance and structure and function of mitochondrial membrane aldehyde dehydrogenase (ALDH) as a final goal, mitochondrial membrane from rat liver was purified using sucrose step gradient (1∼2 M) centrifugation, and then sonicated and ultracentrifuged to obtain pure mitochondrial membrane fraction. The observation of the ALDH activity from this fraction let us know the existance of mitochondrial membrane ALDH. The result that various kinds of detergent (Triton X-100, Digitonin, Lubrol PX, Cholate and Chaps) were examined as a good solubilizer at several different concentrations showed 1% cholate for the best one. In addition to this, the optimum salt (NaCl) concentration and pH for solubilization were 1 M and 9.0, separately. And also, the addition of phospholipid functioned as a stabilizer. The partial purification of ALDH from this cholate-solubilized membrane fraction with selected best conditions was tried using sucrose linear gradient (3∼10%, including 1% cholate and 5 ㎎/㎖ phospholipid) centrifugation and was obtained more 100 times than crude mitochondrial state as a result. This partial purified, cholate-solubilized ALDH was dialyzed to remove the cholate and to make ALDH-incorporated vesicles. ALDH activity from this result was increased about 2 times than before dialysis.
Ti plasmid vector 를 이용한 진핵세포 유전자의 도입에 관한 연구
이희봉,주충노,홍순주,김성완,임창진,김영명 ( Hee Bong Lee,Chung No Joo,Sun Joo Hong,Seong Wan Kim,Chang Jin Lim,Young Myeong Kim ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.1
DNA recombination using plasmid pGA658, a Ti plasmid vector, was tried to study the role of yeast homoserine dehydrogenase after introducing its gene into plant cells. Two recombinant plasmids with opposite orientation of the gene DNA under nos promoter of vector pGA658, pKDB1 and pKDB2, were constructed by the aid of two intermediate vectors, pUC7 and pUC119. Transformed E. coli and Agrobacterium tumefaciens were confirmed by the resistance to appropriate antibiotics and by sizing DNA restriction fragments after plasmid isolation. Assay of homoserine dehydrogenase activity showed that a little increase in its activity was detected in transformed E. coli. but no increase in transformed Agrobacterium tumefaciens, compared with its untransformed strain.
Ti Plamid Vector System 을 이용한 외래 유전자의 도입 : ( 2 ) E . coli thioredoxin 유전자의 배양된 담배세포내 발현
이희봉,주충노,홍순주,김성완,임창진,김영명 ( Hee Bong Lee,Chung No Joo,Soon Joo Hong,Seong Wan Kim,Chang Jin Lim,Young Myeong Kim ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.4
This study was performed to observe the expression of E. coli thioredoxin gene incorporated in tobacco cells. The recombinant DNA used, pKDB3, had been constructed from a Ti plasmid vector pGA658 and a bacterial plasmid pCJF4 harboring E. coli thioredoxin gene, as described in the preceding paper (Lee et al., 1988). The leaf discs of plant (N. tabacum cv Xanthi) were transformed to kanamycin resistance by the cocultivation with Agrobacterium A281 containing plasmid pKDB3. Transformed leaf discs were cultured in MS agar medium with kanamycin for callus induction. Kanamycin-resistant tobacco calli were continuously grown in MS agar medium for shoot induction, and then in MS agar medium for root induction. Expression of E. coli thioredoxin gene in the plant tissue regenerated from transformed tobacco cells was confirmed by DTNB assay. The thioredoxin activity of transformed tobacco cells was much higher (about 9 times) than that of normal tobacco cells. Our results suggest that E. coli thioredoxin gene was successfully incorporated into tobacco cells, and the incorporated bacterial gene could be expressed at a high level.
Ti Plasmid Vector System 을 이용한 외래 유전자의 도입 : ( 1 ) A . tumefaciens 로의 E . coli Thioredonxin 유전자의 도입
이희봉,주충노,홍순주,김성완,임창진,김영명 ( Hee Bong Lee,Chung No Joo,Soon Joo Hong Seong Wan Kim,Chang Jin Lim,Young Myeong Kim ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.4
In this part of study on the incorporation of foreign gene into plant cells, a derivative of Ti plasmid vector (pGA658), containing E. coli thioredoxin gene, was prepared and introduced into Agrobacterium tumefaciens. A recombinant plasmid, pKDB3, was constructed by transferring HindIII-BamHI DNA fragment of pCJF4, including E. coli thioredoxin gene, into HindIII-BgIII restriction sites of plasmid pGA658. By doing this, E. coli thioredoxin gene is expected to express from nos promoter of pGA658 after the incorporation into plant cells. The structure of DNAs isolated from kanamycin-resistant E. coli transformants was convinced by restriction mapping. As a preceding step before incorporation into plant cells, the recombinant plasmid pKDB3 was transformed into A. tumefaciens by freeze-thaw procedure. In Agrobacterium transformants, the expression of E. coli thioredoxin gene was positively observed, and this suggested the stable existence of the E. coli gene.
이희봉,주충노 ( Hee Bong Lee,Chung No Joo ) 생화학분자생물학회 1983 BMB Reports Vol.16 No.2
^(14)C-labelled ginseng saponins from ^(14)C-acetate were prepared using the root slices of panaxginseng C.A. Meyer as an enzyme source and the products were demonstrated to be identical with the saponins extracted from the roots by thin layer chromat ography, high performace liquid chromatography and autoradiography. The radioactivity of the liver was monitored on time course after the feeding of 1 ㎎ of ginseng saponin containing ^(14)-C-labelled saponin. The extract from the liver of rats killed at one hr. after the oral administration of 1 ㎎ of ginseng saponins containing the ^(14)C-saponins was analyzed found that 25 % of the radioactivity recovered was found as undissociated form of the saponin. It semed that the half life of the saponin might be 4∼6 hrs and the concentration of the saponin would be 10^(-5) % level in the liver.
상류(上流) 전통주거(傳統住居) 해남(海南) 녹우당(綠雨堂)의 해석(解釋) -거주자(居住者) 생활(生活)과 농업경영(農業經營)으로-
이희봉,이향미,Lee, Hee-Bong,Lee, Hyang-Mee 한국건축역사학회 2002 건축역사연구 Vol.11 No.1
This study has main purpose to understand and interpret the house Nok-U-dang, an upper class built in Chosun dynasty, not by outsider researcher's view but insider dweller's view. To interpret correctly, dweller's everyday life in the space and form is examined on a microscale beyond the physical space and form of the house, main object of architectural history To understand the present form exactly, the study restore traditional life in past era, 1940s. Main method of restoration is the ethnographic interview, based on cultural anthropology. Like any other upper-class house, the house has been influenced under ruling Confucian ideology in Chosun dynasty: separation of man's and woman's quarters and hierachical arrangement by generation, and worshipping ceremony for ancestor. However, it is by practical management for agricultural production that every court and building of the Noku-Dang can be explained correctly; preparing seed for sowing, tool storing, preparing and serving meal for laborers, making manure, harvesting, threshing grain, storing grain and so on. Precedent studies interpreted the house by the Confucian principle too much and made conclusion of dignity and austerity of ritual: woman's quarter, is closed and serene space. However this study shows that the space is semi-opened and composite space by agricultural works. And the Sarangchae, master's quarter, is located properly at visual center to control every agricultural activity.