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김도훈,양보경,김현경,김나영,정순재,김익수,남재성,이재헌,정대수,Kim, Doh-Hoon,Yang, Bo-Kyung,Kim, Hyeon-Kyoung,Kim, Na-Young,Jeong, Soon-Jae,Kim, Ik-Soo,Nam, Jae-Sung,Lee, Jai-Heon,Chung, Dae-Soo 한국식물생명공학회 2003 식물생명공학회지 Vol.30 No.3
Genetic variation of Perilla germplasms was investigated using RAPD markers. Forty-two Perilla frutescens lines and cultivars collected form locals were subjected to RAPD analysis using 220 primers. Among them only 13 primers showed polymorphic bands and these 13 primers provided a total of 144 bands, consist of 115 polymorphic and 29 monomorphic ones. The polymorphic bands were subjected to phylogenetic analysis using UPGMA and maximum parsimony (MP) methods. In the UPGMA method, similarity coefficiency of 42 Perilla frutescens lines and cultivars ranged from 0 to 0.7842. The dendrogram of 42 lines and cultivars obtained through UPGMA method resulted in two major groups, and the similar clustering pattern was found by MP method, suggesting Perilla germplasms utilized in this study truly can be divided into two major groups. Although the two major groups were consistent roughly with their phenotypes (under of node, weight of 1,000 grains, and oil content), in detail, much inconsistency also was present.
사탕무(Beta vulgaris L.) 의 자엽소 배양에 의한 callus 유기 적색색소 및 단백질 \ulcorner량의 변화
김현경,김도훈,정대수,박현진,Kim, Hyoun-Kyoung,Kim, Doh-Hoon,Chung, Dae-Soo,Park, Hyeon-Jin 한국생명과학회 1997 생명과학회지 Vol.7 No.4
In order to produce betacyanins from sugar beet(Beta vulagris L.) in vitro, callus induction, shoot formation, root formation, betacyanin contents and protein contents determined from callus which was induced cotyledons of sugar beet seedling under an addition of NAA and BAP on the MS medium. The results were summarized as follows; The combination 3.0mg/l NAA and 1.0mg/l BAP treatment showed the most high callus induction rate, betacyanin and protein contents. The combination NAA and BAP treatments were not shoot formation, but BAP treatments showed high root formation rate. But high concentrations of BAP have not shown root formation.
양원태,김경숙,권용삼,김두현,김도훈,Yang, Won Tae,Kim, Kyoung Sook,Kwon, Yong Sham,Kim, Du Hyun,Kim, Doh Hoon 한국식물생명공학회 2016 식물생명공학회지 Vol.43 No.4
본 연구는 육종용을 이용한 기능성 화장품 소재 개발을 목적으로 육종용 추출물의 피부 미백 및 항노화 효능 검정을 위해 수행되었다. 육종용 추출물의 세포 생존율은 B16F10 melanoma 세포를 이용하여 MTT assay를 수행하여 측정한 결과, 육종용 추출물 4 mg/ml 농도에서 90% 이상의 세포 생존율을 보였다. 육종용 추출물을 처리한 B16F10 melanoma 세포를 이용하여 RT-PCR로 유전자 발현을 검정한 결과 tyrosinase, collagenase 및 elastase 유전자의 발현을 육종용 추출물 4 mg/ml 농도에서 각각 80.9%, 37.6%, 70.9% 억제 하였다. 육종용 추출물의 DPPH 라디칼 소거능 측정을 이용한 항산화 활성은 2 ~ 10 mg/ml 농도에서 70.6% ~ 82.6%로 높게 나타났다. 육종용 추출물의 미백과 항노화 효과를 검정하기 위하여 육종용 추출물을 2, 4, 6, 8 및 10 mg/ml 농도로 처리하여 확인하였다. 육종용 추출물의 tyrosinase 저해활성은 66.8% ~ 78.5%, elastase 저해활성은 67.6% ~ 79.3%, collagenase 저해활성은 72.3% ~ 83.6%로 각각 나타났으며, hyaluronidase 저해활성은 65.8% ~ 69.2%로 나타났다. 따라서 육종용은 피부 미백 및 노화방지를 위한 기능성 화장품 소재로 우수한 것으로 평가되며, 세포생존율과 각종 효능을 고려할 때, 육종용 추출물 4 mg/ml 농도가 적정농도로 판단된다. This study assessed the whitening and anti-aging effects of the Cistanche deserticola extract, to develop a cosmetic substance. The cell viability of the Cistanche deserticola extract was evaluated in B16F10 melanoma cells by the MTT (3-(4,5-dimethylthaiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. The cell viability of the extract was determined to be 90% at 4mg/ml concentration. Furthermore, the tyrosinase, collagenase, and elastase mRNA expression level were measured by RT-PCR, using the Cistanche deserticola extract treated B16F10 melanoma cells. At 4 mg/ml concentration, mRNA expression level of tyrosinase, collagenase, and elastase was dramatically decreased to 80.9%, 37.6%, and 70.9%, respectively. The antioxidant activity of the Cistanche deserticola extract was determined by DPPH free radical scavenging. The DPPH free radical scavenging capacities ranged from 70.6% to 82.6%, when evaluated from 2 mg/ml to 10mg/ml concentrations. The effects of whitening and anti-aging of the Cistanche deserticola extracts were examined at 2, 4, 6, 8, and 10 mg/ml concentration. Tyrosinase activities were inhibited from 66.8% to 78.5%, elastase activities were inhibited from 67.6% to 79.3%, collagenase activities were inhibited from 72.3% to 83.6%, and hyaluronidase activities were inhibited from 65.8% to 69.2%, respectively. These data suggest that the Cistanche deserticola extract is effective in whitening and anti-aging; therefore, it is considered to be a functional cosmetic material in cosmetic products.
한국 동해안에서 서식하는 진주담치(Mytilus edulis)의 미토콘드리아 DNA 다형현상
김익수,민병윤,윤명희,김도훈,Kim, Ik-Soo,Min, byung-Yoon,Yoon, Myung-Hee,Kim, Doh-Hoon 한국생명과학회 1999 생명과학회지 Vol.9 No.3
Mitochondrial DNA (mtDNA) polymorphism of the blue mussel (Mytilus edulis) species complex sampled from the east coast of Korean was studied using a partial sequence of COIII gene (336 bp). Samples obtained from three localities on the east coast of Korea revealed four haplotypes with two clearly differentiated mitochondrial clades (termed clades B and E), separated by 4.2% of minimum sequence divergence. This pattern indicates no difference between east and south coasts of Korea. According to population genetic theory on evolutionary characteristics of mtDNA, we concluded that mtDNA introgression from M. edulis to M. gallprovincialis might be a source for mtDNA polymorphism found in mussels on the east coast of Korea.
백선의 형태적 특성, 항균 효과 및 분자 세포학적 분석
정대수,조창우,김경미,허지은,김수영,정영수,김도훈,방재욱,최정,이재헌,Chung, Dae-Soo,Cho, Chang-Woo,Kim, Kyung-Mee,Heo, Jee-Eun,Kim, Soo-Young,Chung, Young-Soo,Kim, Doh-Hoon,Bang, Jae-Wook,Cui, Zheng,Lee, Jai-Heon 한국생명과학회 2006 생명과학회지 Vol.16 No.7
The importance of wild plant resources along with the development of high biotecyhnology is highlighted for exploitation of new materials which can make the added value. The goal of this study is providing fundamental data bases for developing new materials through the analyses of morphological characteristics, antibiotics and molecular cytogenetics in Baegseon (Dictamnus dasycarpus Turcz.). Baegseon has several characteristics; there are two types of flower color, pink and white, the seed germination starts about February 20th, the maximum flowering season is around May 17th in southern Korea, the growth period is about 60 days. The number of chromosomes are 2n=2x=36, the size of chromosomes in metaphase is $4.2{\sim}8.1{\mu}m$. The amount of 2C nuclear DNA is 1.93pg, and there is no variation of genome size amoung varieties. The extraction juice of baegseon young roots has the excellent antibiotic activity against the mold (Botrytis cinerea).
이혜영,이기정,전은희,신상현,이재헌,김도훈,정대수,정용모,조용조,김정국,정영수,Lee, Hye-Young,Lee, Ki-Jung,Jeon, Eun-Hee,Shin, Sang-Hyun,Lee, Jai-Heon,Kim, Doh-Hoon,Chung, Dae-Soo,Chung, Yong-Mo,Cho, Yong-Cho,Kim, Jeong-Kook,Chung, You 한국식물생명공학회 2006 식물생명공학회지 Vol.33 No.1
본 연구에서는 거베라 형질전환 기초기술개발을 위한 연구를 수행하였다. 경남원예화훼시험장에서 육성된 국내 거베라 18품종 중, Agrobacferium 감염에 순응적인 품종을 선발하기 위하여 18개의 국내 거베라 품종을 스크린하여 011, 016, GM023 등 12개의 순응형 계통을 스크린 하였으며 효율적인 형질전환을 위한 제균 항생제로는 cefotaxime이 선택되었다. 더 많은 캘러스 유도를 위한 전처리 배지의 호르몬 농도는 BA 2 ppm, Zeatin 2 ppm, IAA 0.2 ppm (2 x media)이 효율적이었고 재분화 배지의 호르몬 농도도 전처리 배지 호르몬의 농도와 동일하게 BA 2 ppm, Zeatin 2 ppm, IAA 0.2 ppm (2 x media)에서 효율적이었으나 GUS 발현에는 다른 차이가 없었다. 고효율 형질전환 체계 확립을 위하여 네 가지 다른 전처리 기간 처리와 두 가지 다른 Agrobacterium tumerfaciens 사용, 두 가지 다른 공배양 기간을 처리한 결과 전처리 하지 않은 엽병과 엽신을 LBA4404 Agrobacterium tumerfaciens을 사용하여 5일 간 공배양 기간을 가졌을 때와 dipping 처리하여 접종한 엽병과 엽신에서 높은 GUS 발현빈도를 나타냈다. Gerber (Gerbera hybrida) is a valuable ornamental species grown as a potted plant and cut flowers. However, genetic variability within the gerbera genus is very limited. So it is absolutely needed to introduce and widen genetic resources into gerbera lines by genetic transformation. For the purpose, 18 Korean gerbera lines were screened to establish Agrobacterium-mediated genetic transformation procedure. In an experiment to select Korean gerbera lines which are amenable to Agrobacterium-inoculation, 12 lines turned out to be positive in Agrobacterium-inoculation. More callus were produced from BA 2ppm, Zeatin 2ppm, IAA 0.2ppm in pre-culture and regeneration medium (2X media) but there was no difference in the frequency of GUS expression rate. In another experiment to find out optimal condition for highly efficient Agrobacterium-inoculation, petiole and leaf explants have been treated with four different pre-culture periods, two different co-culture periods and two different Agrobacterium strains. As a result, high GUS expression has been showed from petiole and leaf explants treated no pre-culture period with LBA4404 Agrobacterium tumerfaciens, 5 day co-culture period and dipping treatment.
국내 콩(Glycine max) 품종 형질전환 초기조건 확립
이기정,서진경,이혜영,전은희,신상현,이재헌,김도훈,고종민,한원영,백인열,오병준,정영수,Lee Ki-Jung,Seo Jen-Kyung,Lee Hye-Young,Jeon Eun-Hee,Shin Sang-Hyun,Lee Jai-Heon,Kim Doh-Hoon,Ko Jong-Min,Hahn Won Young,Baek In-Youl,Oh Boung-Jun,Chung 한국생명과학회 2006 생명과학회지 Vol.16 No.2
경제적으로 중요한 식용작물인 콩(Glycine max)의 초기형 질전환 효율증대를 위하여 연구를 수행하였다. 높은 초기형 질전환 조건 확립을 위하여 기초실험으로 Agrobacterium 감 염에 순응적인 국내 콩 품종을 스크린 하였으며, 적정한 agar 농도와 선발항생제 hygromycin 농도 규명, 상처방법, explant의 치상방향, 호르몬 전처리, 액체배지 선발 등 형질 전환 초기효율에 미치는 영향을 transient GUS 분석을 통하여 알아보았다. Agrobacterium 감염에 순응적인 품종을 선발하기 위하여 32개의 국내 콩 장려품종을 스크린한 결과, 일품검정콩, 은하콩, 만리콩, 대원콩 등 14개의 순응형 품종을 선발하였으며, 효율적인 제균제로는 cefotaxime이, 효율적인 선발항생제로서는 hygromycin이 선택되었다. Hygromycin 농도는 10-15 ppm이 효율적이었고, agar 농도 0.6-0.8%, explant의 치상방향은 향배축(adaxial side)을 down하여 치상하였을 때가 높은 GUS ($\beta$-glucuronidase)발현빈도를 나타냈고 상처방법은 scalpel보다. 동양침을 사용하였을 때 높게 나타났다. 형질전환 체계확립을 위하여 호르몬 전처리를 한 결과, BA 5 ppm 과 10 ppm을 처리하였을 때 후기 선발에서 높은 생존율을 보였고, 전처리 후에 액체배지에서 선발을 하였을 때 비형질 전환체(escape)의 빈도를 크게 낮출 수 있었다. 또한 여러 조건의 결과를 비교하여 볼 때, 국내 콩 품종 중에서 형질전환을 위한 재료로써는 은하콩이 가장 적합한 것으로 생각된다. In order to establish highly efficient gene transfer condition at early stage of soybean transformation, various experiments were performed and compared their efficiencies by transient GUS analysis; those conditions are genotype determination of Korean soybean cultivars for amenability to Agro-infection, appropriate agar and selective agent concentration, orientation of explant placement, hormone pre-culture, and liquid selection condition. In the genotype screen of Korean soybean varieties, 14 amenable genotypes were selected. For efficient Agrobacterium washing, cefotaxime was chosen and hygromycin at the concentration of 10 and 15 ppm was used as selection agent in the media. Agar concentration was slightly better in 0.6% and 0.8% for both shoot and callus formation, and explant placement with adaxial side down showed high frequency of GUS expression. For wounding treatment, oriental needle was efficient than scalpel for shoot formation and gene transfer. To increase the frequency of gene transfer, hormone pre-treatment was applied. BA at the concentration of 5 and 10 ppm resulted in better survival at the late stage of selection in shoot elongation media. Selection in liquid media after hormone pre-treatment seemed to be effective to remove the escaped non-transformants at early stage of procedure. Considering the results obtained, Eunhakong could be the first choice as a material for soybean transformation among Korean soybean genotypes.