TAR cloning 법에 의한 인간 및 마우스의 상동성 HPRT 유전자의 분리

도은주,김재우,정정남,박인호,임선희,Do, Eun-Ju,Kim, Jae-Woo,Chung, Chung-Nam,Park, In-Ho,Leem, Sun-Hee 한국생명과학회 2006 생명과학회지 Vol.16 No.6

TAR (Transformation-Associated Recombination) cloning법은 복잡한 고등생물의 게놈으로부터 유전자나 특정 염색체 부위를 선별적 분리를 가능하게 한다. 이 방법은 목적으로 하는 염색체 부위의 주변에 존재하는 비교적 짧은 게놈 염기서열에 대한 정보를 필요로 한다. 이 기술은 출아효모의 spheroplasts 형질전환 동안 목적 유전자를 포함한 게놈 DNA와 그 유전자의 5' 또는 3' 말단 서열 (hook)을 포함하고 있는 TAR vector 사이에 일어나는 상동성 재조합에 의해 이루어진다. 본 연구에서는 TAR cloning 법을 상동성 유전자의 분리에 사용할 수 있는가를 조사하기 위해, 연간과 마우스 게놈의 HPRT 유전자를 선택하였다. 그 결과, 인간과 마우스의 게놈으로부터의 HPRT 유전자의 분리 빈도는 TAR vector로서 hHPRT hook 혹은 mHPRT hook을 사용한 경우에 거의 동일하게 나타났다. 또한 mHPRT 유전자의 gap 부분의 염기서열을 결정하여, 이 부분에 염기서열의 불안정의 요인이 되는 비정상적 특성을 발견하였다. 결론적으로 TAR cloning법을 이용하여 다른 이종 간의 게놈으로부터 상동성 유전자 즉 orthologue의 분리가 가능하였다. 더욱이 TAR cloning 시스템을 이용하여 고등동물 게놈 상에 남아있는 gap 부분을 메움으로서 고등동물의 모든 유전자들의 확인이 가속화될 수 있을 것으로 사료된다. The transformation-associated recombination (TAR) cloning technique allows selective isolation of chromosome regions or genes from complex genome. The procedure requires knowledge of relatively small genomic sequences that reside adjacent to the chromosome region of interest. This method involves homologous recombination during spheroplast transformation between genomic DNA and a TAR vector that has 5' and 3' gene targeting sequences (hooks). To examine whether TAR cloning can be applied to the isolation of gene homologues, we chose the HPRT genes from human and mouse genome. As results, the yield of positive clones for HPRT gene from human and mouse genome when using a TAR vector containing mHPRT hook or hHPRT hook was almost same level. Analysis of the gap regions in mHPRT revealed that they contain abnormalities that could result in instability of the sequences. In conclusion, we were able to use the TAR cloning technology to isolate gene homologue (orthologue) from nonidentical genome. Moreover, the use of the TAR cloning system may accelerate work on closing the remaining gaps in mammalian genome to achieve the goal of annotation of all mammalian genes.

Cyclodextrin Glucanotransfease 고생산 호알칼리성 세균의 탐색과 분비 효소의 특성

도은주,박종부,이용현 한국산업미생물학회 1993 한국미생물·생명공학회지 Vol.21 No.2

An alkalophilic microorganism for overproduction of cyclodextrin glucanotransferase(CGTase) was newly isolated from hot-water spring soil, and identified as Bacillus firmus var, alkalophilus H609. The strain maintained stability during preservation and cultivation for the enzyme production, and produced significant amount of CGTase corresponding to the volumetric activity of 75 units/mL at 37℃, initial pH of 11.2, and after 40 hours. The strain excreted several different proteins showing CGTase activity that catalyzed the formation of mainly β- and γ-type cyclodextrin (ratio of 7:1) from soluble starch without accumulation of α-type. Other enzymatic properties were also investigated.

한약재 추출물 함유 앰플의 주름개선 효능에 관한 기초 임상연구

도은주,이진상,박현진,하일도,김영철,김미려,Do, Eun-Ju,Lee, Jin-Sang,Park, Hyun-Jin,Ha, Il-Do,Kim, Young-Chul,Kim, Mi-Ryeo 대한한의학방제학회 2009 大韓韓醫學方劑學會誌 Vol.17 No.2

Objectives : This study was performed to evaluate the efficacy of ampule containing herbal extract(Ulmus davidiana, Ginkgo biloba, Perilla ocymoides, Morus alba, Glycyrrhiza uralensis (licorice), and Angelica gigas) on skin wrinkles in adult women with facial wrinkles. Methods : A total of 12 women, 30 to 46 years of age, with wrinkles in the corner of eyes, applied ampule containing herbal extract twice daily for up to 12 weeks. Silicon replicas of right crow's feet area and antecubital fossa were taken before use of test product and at 4, 8 and 12 weeks. The replicas were analyzed by optical profilometry with Skin Visiometer SV600. The wrinkle and roughness parameters, R1, R2, R3, R4 and R5 were calculated and statistically analyzed. In addition, a subjective evaluation of product efficacy was conducted by patient's assessment. Results : The mean values of all the skin roughness parameters were decreased at 4-week, and decreased significantly at 8 and 12-week except R4 at 12-week after test product use. The subjective evaluation of wrinkle by patient's assessment was also improved. However, one patient noted stiffness of face after use of test product. Conclusions : These study suggested that the application of ampule containing herbal extract twice daily for 12 weeks may effectively improve the facial wrinkles without severe side effect.

수술실 내의 대기시간 단축

도은주 한국의료QA학회 2005 가을학술대회 Vol.2005 No.2

경옥고가미방 추출물이 생쥐의 양모 및 발모 관련 단백 발현에 미치는 영향

도은주 ( Eun Ju Do ),황미열 ( Mee Yul Hwang ),김승연 ( Seung Yeon Kim ),이진상 ( Jin Sang Lee ),양대석 ( Dae Seok Yang ),양재하 ( Chae Ha Yang ),김미려 ( Mi Ryeo Kim ) 대한본초학회 2011 大韓本草學會誌 Vol.26 No.4

Objective: Present study was carried out to investigate the effect of Gyungohkgo-gamibang extract on hair growth and protein expression in an alopecia model of C57BL/6 mice. Methods: Mice were divided into 3 experimental groups including normal (vehicle), Gyungohkgo-gamibang extract (YNS-10) and 5% minoxidil-treated group. The test materials were daily applied with 0.1 ml per mouse on shaved dorsal skin for 3 weeks. The hair growth was monitored by photograph at 5, 10, 15, 21 days after topical application. Then the changes of hair density and hair thickness in the hair-removed area were evaluated by phototrichogram using folliscope. Also the expression level of growth factors related to hair growth was measured by western blotting. Results: Application of minoxidil or YNS-10 stimulated the hair growth compared to vehicle treatment. Therefore hair density of minoxidil or YNS-10 application was increased about 200% and 210% more than in vehicle application on 14 day, respectively. And hair thickness of both minoxidil group and YNS-10 group was increased about 220% and 210 % more than in vehicle spreading on 14 day, respectively. Futhermore the protein expression of IGF-1 and VEGF were significantly up-regulated on 7 day in YNS-10 and minoxidil-spreaded group compared to vehicle-applied group. Conclusion: These data suggest that YNS-10 has potent stimulating activity on hair growth in C57BL/6 mice and potential usefulness as ingredients of hair tonic and hairrestore.

여주 추출물을 함유한 한방화장품의 주름 및 미백 개선에 관한 임상적 연구

정호준,도은주,이진상,박현진,오규석,김미려,하일도,정용하,지선영,Jung, Ho-Jun,Do, Eun-Ju,Lee, Jin-Sang,Park, Hyun-Jin,Oh, Gyue-Seok,Kim, Mi-Ryeo,Ha, Il-Do,Jeong, Yong-Ha,Jee, Seon-Young 대한한방안이비인후피부과학회 2011 한방안이비인후피부과학회지 Vol.24 No.2

Objective : The purpose of this clinical research was to investigate the effects of herbal cosmetics containing Momordica charntia L. extracts on the anti-wrinkle and whitening effects. Methods : A total of 25 subjects who visited Daegu Hanny Oriental Medical Center from November 1st, 2010 to December 31th, 2010 were included in this study. In this study, we observed change of R1, R2, R3, R4, R5 for using products, change of average melanin content and pigmental area, analysis results of subjective wrinkle improvement, Analysis results of subjective whitening improvement. Statistical analysis was performed by using paired t-test and wilcoxon signed ranks test. Statistical significance was achieved if the probability was less than 5%(p<0.05) Results : Statistically, herbal cosmetics containing Momordica charntia L. extracts showed siginificant effect on change of R1, R2, R3, R4, R5 for using products, change of average melanin content and pigmental area, analysis results of subjective wrinkle improvement, Analysis results of subjective whitening improvement(p<0.05). And satisfaction after using herbal cosmetics was near good. Conclusions : Considering the above results, we have concluded that herbal cosmetics containing Momordica charntia L. extracts have the remarkable effects on the anti-wrinkle and whitening effects.

구미전자산업의 연계체계와 공간적 특성 : Electronics Industry in Kumi, South Korea

이철우,도은주 慶北大學校 社會科學大學 1996 社會科學 Vol.8 No.1

이 논문은 구미전자 산업을 사례로 최근 공업지리학의 주요 연구주제의 하나인 기업조직 측면에서의 연계패턴과 그 공간적 특성을 현지조사를 중심으로 연구하고자 하였다. 구미전자산업의 경우 다공장형 기업의 비중이 높음에도 불구하고 전 구성요소에 걸쳐 기업조직내의 연계보다는 타 독립기업간의 연계가 강한 ‘사회적 분업형’의 성격이 강하며 서비스 연계를 제외하고는 상업기능과의 연계보다 제조업체간의 연계가 강하다. 공간적으로는 국지적 고객연계와 비국지적 공급자 연계가 상대적으로 강하여 연계체계상의 고객기업이 선도산업으로서 지역경제에 미치는 영향은 절대적이다. 물론 이러한 선도기업에 부품공급과 하청관계를 중심으로한 지역내 신설기업이 늘어남에 따라 국지적 공급자 연계가 강화되는 경향을 보이고 있다. 그럼에도 불구하고 생산자 서비스를 중심으로한 서비스 부문의 역내연계가 매우 미약하여 범위의 경제를 추구함에 있어서 제약이 크다. The purpose of this study was to analyze the linkage pattern and its spatial characteristics. The data were come from mail survey and face-to-face interview with the electronics firms in Kumi, South Korea. The social division of labor among businesses was easily found; while, there is the great portion of a multi-plant business. Most research discovered that a multi-plant business inclined to maintain a intra firm linkage within the business rather than a linkage with other businesses. The linkage among manufacturing firms is stronger than the linkage between manufacturing firm and commercial business in a purchasing linkage, a subcontract linkage and a marketing linkage except a service linkage. In the system of linkage, a local customer linkage and a non-local supplier linkage were strenghten; while, a local supplier linkage and a non-local customer linkage were relatively weak. In the system of linkage, customer firms is leading industry as the result of that customer firms strongly impacted on regional economics. The local supplier linkage intended to be strong as the result of the creation of the number of new firms that supply the parts to the leading industry. Most of the new firms are small and medium enterprises. A vertical linkage was established between a leading industry and a small and medium enterprises. The major fact of the establishment of local linkage in Kumi was the degree of quality on goods and service as well as the good accessibility in terms of communication, time and/or monetary cost of transportation. Most research ignored the degree of quality of goods and service in establishing local linkages. It was recommended that Kumi Electronic Industry should attempt to reinforce local supplier linkage in basic materials, machinery and producer services; and change the linkage system from a vertical linkage between a leading industry and a small and medium enterprises to a horizontal linkage.

Target DNA 염기서열 내에 존재하는 비상동성 간격이 상동성재조합을 이용한 클로닝 빈도에 미치는 영향

김재우,도은주,윤세련,정윤희,윤영호,임선희,선우양일,박인호,Kim Jae-Woo,Do Eun-Ju,Yoon Se-Lyun,Jeong Yun-Hee,Yoon Young-Ho,Leem Sun-Hee,Sunwoo Yangil,Park In-Ho 한국미생물학회 2005 미생물학회지 Vol.41 No.4

Transformation-associated recombination (TAR) 클로닝 법은 복잡한 게놈으로부터 염색체 내의 특정부위나 유전자를 선택적으로 분리할 수 있다. 이 방법은 목적 유전자에 근접한 작은 게놈DNA 염기서열 정보를 필요로 한다. 이 기술은 효모의 spheroplast transformation을 시키는 동안 목적으로 하는 유전자의 5' 또는 3' 서열을 포함하고 있는 TAR vector와 게놈DNA사이에서 일어나는 상동성재조합에 의해 이루어진다. 본 연구에서는 plasmid 모델시스템을 이용하여 target hooks 내에 존재하는 비상동성 염기서 열이 상동성재조합에 미치는 영향을 조사하였다. plasmid에 존재하는HIS3유전자와 변형시킨 his3-TRP1-his3 단편 사이의 상동성재조합의 효율은 $Ura^+$ 형질전환체의 형질분석에 의해 이루어졌다. $Ura^+$ 형질전환체의 수는 7종류의 서로 달리 변형된 his3-TRP1-his3 단편들을 사용하였을 매 거의 동일하게 나타났다. 그러나 $Trp^+His^+$ positive recombinants의 빈도는 변형된 his3-TRP1-his3 단편 내에 비상동성 영역에 부정확한 간격을 지닐 때 현저한 감소를 나타내었다. 이러한 결과로서, 부정확한 간격이 target hook과 substrate DNA 사이에 일어나는 상동성재조합을 방해하는 것으로 사료된다. 그러므로 이종간의 상동유전자를 클로닝 할 때에는 target hook내의 비상동성 염기서열이 존재한다면 이것이 정확한 간격을 지니는지 여부를 중요란 요인으로 고려해야 한다. Transformation-Associated Recombination (TAR) cloning technique allows selective isolation of chromosomal regions and genes from complex genomes. The procedure requires knowledge of relatively small genomic sequences that reside adjacent to the chromosomal region of interest. This technique involves homologous recombination during yeast spheroplast transformation between genomic DNA and a TAR vector that has 5' and 3' gene targeting sequences. In this study, we examined the effect of non-homologous spacing sequence in target hooks on homologous recombination using a plasmid model system. The efficiency of homologous recombination between the modified his3-TRP1-his3 fragments and HlS3 gene on plasmid were analyzed by the characterization of $Ura^+$ transformants. The numbers of $Ura^+$ transformant showed same level when seven different modified his3-TRP1-his3 fragments were used. But the percentage of positive recombinants. $Trp^+His^-$, dramatically decreased when used the modified his3-TRP1-his3 fragments contained incorrect spacing of nonhomologous region. As a result, we suggest that incorrect spacing inhibits the homologous recombination between target hook and substrate DNA. Therefore, we should consider the correct spacing in target hook when the target hook are used for cloning of orthologue gene.

알코올을 투여한 흰쥐의 알코올 대사에 미치는 한약재 혼합추출물의 영향

정미진,김소영,도은주,윤종국,김대익,한경수,김미려 대한본초학회 2019 大韓本草學會誌 Vol.34 No.1

Objectives : Alcohol hangover is a common phenomenon which basically occurs after heavy drinking. Moreover, heavy alcohol consumption leads to acute and chronic diseases. We investigated the effect of herbal mixture (SJ) on alcohol metabolism in serum or/and liver. Methods : 5W-old Sprague Dawley rats were used for the study. To overnight fasted rats, 0.9% saline or SJ extract was administrated per os before alcohol treatment. Then, 40% alcohol was orally administrated to all rats in 30 mins. Ethanol, acetaldehyde concentrations, alcohol dehydrogenase (ADH), acetaldehyde dehydrogenase (ALDH) activities were measured by assay kits. Aspartate aminotransferase (AST) and alanine transaminase (ALT) were measured by analyzer. Results : Ethanol and acetaldehyde concentrations lowered in SJ groups compared with CON group. Especially, acetaldehyde concentration significantly decreased in SJ groups compared with CON group. AST and ALT levels tended to increase in SJ groups compared with CON group but there was no significant difference between CON group and SJ groups. ADH activity in serum was higher in SJ groups than CON group but no significant difference in liver. ALDH activity in both serum and liver showed significantly increased in SJ groups compared with CON group. Conclusions : Treatment of SJ extract showed not only reducing concentration of alcohol and acetaldehyde but also increasing activities of ADH and ALDH. These results suggest SJ may influence in alcohol metabolism via control of metabolic enzyme activities and metabolite. Therefore, SJ, herbal mixture, might have a function of preventing hangover after drinking alcohol.

아토피 피부염 환자에서 상황버섯 추출물을 함유한 한방화장품의 임상적 연구

정호준,황보민,도은주,석장미,김미려,김영훈,도국배,이창언,지선영,Jung, Ho-Jun,HwangBo, Min,Do, Eun-Ju,Suk, Jang-Mi,Kim, Mi-Ryeo,Kim, Young-Hun,Do, Kook-Bae,Lee, Chang-Eon,Jee, Seon-Young 대한한방안이비인후피부과학회 2010 한방안이비인후피부과학회지 Vol.23 No.3

Objective : The purpose of this clinical research was to investigate the effects of herbal cosmetics containing Phellinus linteus extracts on patients with atopic dermatitis. Methods : A total of 35 patients who visited Daegu Hanny Oriental Medical Center from November 4th, 2008 to December 28th, 2008 were included in this study. In this study, we observed transepidermal water loss, skin moisture content, modified scorad index of atopic dermatitis and pruritic degree. Statistical analysis was performed by using paired t-test and wilcoxon signed ranks test. Statistical significance was achieved if the probability was less than 5%(p<0.05). Results : Statistically, herbal cosmetics containing Phellinus linteus extracts showed siginificant effect on transepidermal water loss, skin moisture content, modified scorad index of atopic dermatitis and pruritic degree(p<0.05). And satisfaction after using herbal cosmetics was near good. Conclusions : Considering the above results, we have concluded that herbal cosmetics containing Phellinus linteus extracts have the remarkable effects on atopic dermatitis.