새로운 선발 마커 D-아미노산 산화효소 유전자를 이용한 식물 형질전환

임선형,우희종,이시명,진용문,조현석,Lim, Sun-Hyung,Woo, Hee-Jong,Lee, Si-Myung,Jin, Yong-Moon,Cho, Hyun-Suk 한국식물생명공학회 2007 식물생명공학회지 Vol.34 No.1

Though higher plants car not metabolize D-amino acid, many prokaryotes and eukaryotes have the D-amino acid metabolism. Therefore, we transformed tobacco plants with D-amino acid oxidase (DAO), which can metabolize D-amino acid, and confirmed that transgenic tobacco plants might metabolize D-amino acid. Transgenic tobacco plants were survived a high concentration of D-serine, however non-transgenic plants were not grown on D-serine medium. From Southern and Northern blot analysis, transgenic tobacco plants selected on D-serine medium were confirmed by insert and expression of transgene. $T_{1}$ tobacco seeds derived $T_{0}$ tobacco plants selfing were grown on D-serine medium and showed normal phenotype compared to wild tobacco plants. Transgenic tobacco plants displayed the metabolic capability of D-serine. Therefore, we suggested that DAO is useful selectable marker gene for plant transformation.

환경위해성 평가를 위한 해충저항성 배추의 분자생물학적 특성 검정 및 계통 특이 마커 개발

임선형,조현석,김나영,이시명,우희종,신공식,진용문 한국식물생명공학회 2007 JOURNAL OF PLANT BIOTECHNOLOGY Vol.34 No.4

Commercialization of genetically modified (GM) plants will be required the assessment of risks associated with the release of GM plants that should include a detailed risk assessment of their impacts in human health and the environment. Prior to GM plant release, applicants should provide the information on GM crops for approval. We carried out this study to provide the molecular data for risk assessment of the GM Chinese cabbage plants with insect-resistance gene, modified CryIAc, which we obtained by Agrobacterium- transformation. From the molecular analysis with GM Chinese cabbage, we confirmed the transgene copy number and stability, the expression of the transgene, and integration region sequences between the transgene and the Chinese cabbage genome. Based on the unique integration DNA sequences, we designed specific primer set to detect GM Chinese cabbage and set up the GM cabbage detection method by qualitative PCR analysis. Qualitative analysis with GM Chinese cabbage progenies analysis was revealed the same as the result of herbicide treatment. Our results provided the molecular data for risk assessment analysis of GM Chinese cabbage and demonstrated that the primer set proposed could be useful to detect GM Chinese cabbage.

해충저항성 GM 배추에서 T-DNA와 식물체 게놈의 인접 부위 분석

임선형,조현석,박승혜,김정환,김나영,원소윤,이시명,신공식,우희종,김동헌 한국식물생명공학회 2008 식물생명공학회지 Vol.35 No.2

The Agrobacterium-mediated transformation has been successfully used method to introduce foreign genes into some monocotyledonous as well as a large number of dicotyledonous plants genome. We developed transgenic Chinese cabbage plants with insect-resistance gene, modified CryIAc, by Agrobacterium- transformation and confirmed transgene copy number by Southern blot analysis. We confirmed that twenty-nine out of 46 transgenic Chinese cabbage plants have single copy of CryIAc. To obtain the sequences information on the transferred DNA (T-DNA) integration into plant genome, we analyzed left border (LB) flanking sequences by genome walking (GW) PCR method. Out of 46 transgenic Chinese cabbage plants examined, 37 carried the vector backbone sequences. This result indicates that the transfer of the vector backbone from the binary vectors resulted mainly from inefficient termination of LB site. Analysis of T-DNA LB flanking region of 9 transgenic Chinese cabbage plants without vector backbone revealed that all LB ends were not conserved and nucleotides up to 36bp from the LB cleavage site were deleted.

Marker development for the identification of rice seed color

임선형,하선화 한국식물생명공학회 2013 Plant biotechnology reports Vol.7 No.3

Pigmented traits in rice seeds are regarded asimportant breeding goals for crop improvement. Markerassistedselection is very helpful when screening for targetseed color traits in the early stages of plant development. Among the genes involved in the biosynthesis of anthocyaninsand proanthocyanins (PAs) that are candidates formarker development, we examined the expression of fivegenes encoding CHS, CHI, F3H, DFR, and ANS in theseeds of non-pigmented white and pigmented black and redrice cultivars. The transcript levels of all these genes exceptfor CHI are higher in pigmented rice than in non-pigmentedrice. Sequence variations in these biosyntheticgenes revealed that the DFR gene harbors a single nucleotidesubstitution that generates a premature stop codon inwhite rice. Additional sequence variations in two regulatorygenes, OSB1 and Rc, were also compared among thesame cultivars. The sequence of the OSB1 gene in blackrice was found to differ from that in red and white rice. Thesequence of the Rc gene in red rice also differed from thatin white and black rice. Based on these variations, wedeveloped two CAPS markers for DFR and OSB1 genesand an Indel marker for the Rc gene. The combined use ofthese three markers could discriminate rice seeds harboringwhite, black and red color. We validated the usefulness ofthese markers in 34 rice cultivars. Hence, the combinedapplication of our three new markers may have utility toscreen the seed color prior to seed setting in rice breedingprograms.

유전자총을 이용한 잔디 형질전환 체계 확립

임선형,강병철,남궁용,신홍균 한국잔디학회 2001 한국잔디학회지 Vol.15 No.1

Callus formation and plant regeneration from the seeds of zoysiagrass cv. Zenith was tested on MS basal medium supplemented with various concentrations of 2,4-dichlorophenoxyacetic acid(2,4-D) and of several cytokinins. A concentration of 1mg/L 2,4-D on medium stimulated callus formation. In the presence of 5mg/L 2,4-D, addition of 1mg/L kinetin significantly enhanced callus formation and plant regeneration over 2,4-D alone. To transfer foreign DNA into turfgrass, parameters for the bombardment of embryogenic callus with the particle bombardment were partially optimized using transient expression assay of a $chimeric \beta$-glucuronidase(GUS) gene driven by the CaMV 35S promoter. GUS gene was strongly expressed at helium pressure 1,100 psi and 6~9cm target distance.

유전자총을 이용한 잔디 형질전환 체계 확립 ( Establishment of Transformation Systems of Zoysiagrass by Particle Bombardment )

임선형,강병철,남궁용,신홍균 한국잡초학회·한국잔디학회 2001 Weed & Turfgrass Science Vol.15 No.1

환경위해성 평가를 위한 해충저항성 배추의 분자생물학적 특성 검정 및 계통 특이 마커 캐발

임선형,김나영,이시명,우희종,신공식,진용문,조현석,Lim, Sun-Hyung,Kim, Na-Young,Lee, Si-Myung,Woo, Hee-Jong,Shin, Kong-Sik,Jin, Yong-Moon,Cho, Hyun-Suk 한국식물생명공학회 2007 식물생명공학회지 Vol.34 No.4

유전자변형 작물의 상업화를 위해서는 유전자변형 작물이 식품으로서의 안전성과 환경에 미치는 영향에 관한 평가가 이루어져야한다. 이를 위해 개발자는 유전자변형 작물의 방출이전에 유전자변형 작물 개발에 관한 정보를 제출해야만 한다. 본 연구는 유전자변형 작물의 환경 위해성 평가를 위한 분자생물학적 자료를 제공하고자 수행하였다. 아그로 박테리움 형질전환법을 통하여 해충저항성 CryIAc 유전자가 도입된 배추 형질전환체를 획득한 후, 분자생물학적인 분석을 통하여 유전자의 copy수, 안정성, 식물체내에서의 발현을 확인하였고, T-DNA의 배추게놈내의 인접서열을 분석하였다. T-DNA의 게놈내 삽입 인접서열을 바탕으로 유전자변형 배추를 동정할 수 있는 프라이머를 제작하였고, 이를 이용한 검정방법을 수립하였다. 계통 특이 프라이머를 이용한 해충저항성 배추 후대의 PCR 분석결과와 제초체 처리결과가 서로 일치하였다. 본 연구 결과로 환경위해성 평가를 위한 해충저항성 배추의 분자생물학적인 자료를 획득하였으며, 개발된 프라이머는 해충저항성 배추의 검출을 위하여 유용하게 사용할 수 있음을 확인하였다. Commercialization of genetically modified (GM) plants will be required the assessment of risks associated with the release of GM plants that should include a detailed risk assessment of their impacts in human health and the environment. Prior to GM plant release, applicants should provide the information on GM crops for approval. We carried out this study to provide the molecular data for risk assessment of the GM Chinese cabbage plants with insect-resistance gene, modified CryIAc, which we obtained by Agrobacterium-transformation. From the molecular analysis with GM Chinese cabbage, we confirmed the transgene copy number and stability, the expression of the transgene, and integration region sequences between the transgene and the Chinese cabbage genome. Based on the unique integration DNA sequences, we designed specific primer set to detect GM Chinese cabbage and set up the GM cabbage detection method by qualitative PCR analysis. Qualitative analysis with GM Chinese cabbage progenies analysis was revealed the same as the result of herbicide treatment. Our results provided the molecular data for risk assessment analysis of GM Chinese cabbage and demonstrated that the primer set proposed could be useful to detect GM Chinese cabbage.

식물에서 shikimate 및 방향족 아미노산 생합성 경로와 이의 대사공학적 응용

임선형,박상규,하선화,최민지,김다혜,이종렬,김영미 한국식물생명공학회 2015 식물생명공학회지 Vol.42 No.3

The aromatic amino acids, which are composed of ʟ-phenylalanine, ʟ-tyrosine and ʟ-tryptophan, are general components of protein synthesis as well as precursors for a wide range of secondary metabolites. These aromatic amino acids-derived compounds play important roles as ingredients of diverse phenolics including pigments and cell walls, and hormones like auxin and salicylic acid in plants. Moreover, they also serve as the natural products of alkaloids and glucosinolates, which have a high potential to promote human health and nutrition. The biosynthetic pathways of aromatic amino acids share a chorismate, the common intermediate, which is originated from shikimate pathway. Then, tryptophan is synthesized via anthranilate and the other phenylalanine and tyrosine are synthesized via prephenate, as intermediates. This review reports recent studies about all the enzymatic steps involved in aromatic amino acid biosynthetic pathways and their gene regulation on transcriptional/posttranscriptional levels. Furthermore, results of metabolic engineering are introduced as efforts to improve the production of the aromatic amino acids-derived secondary metabolites in plants.

30-40대 남성의 생활습관과 피부관리 행동이 화장품 구매형태에 미치는 영향

임선형,김성희 대한피부미용학회 2015 대한피부미용학회지 Vol.13 No.4

This study conducted a survey of men in 30s-40s to discuss how the everyday habits and skincare behaviors of men in 30s-40s affect their purchase of cosmetic products. The following summarizes the findings of this study: the most influential factors were the ‘financial properties’, followed by ‘essential properties’, ‘non-essential properties’, and ‘practical properties’. First, based on the demographics, there were significant differences in the financial properties and non-essential properties of purchase of cosmetic products by age, while there was a significant difference in the practical properties by education. Second, significant differences in the financial properties, essential properties, and non-essential properties of purchase of cosmetic products by exercise, but there was no significant difference in the practical properties. Third, there were significant differences in the financial properties, essential properties, and non-essential properties of purchase of cosmetic products by the use of exfoliator, use of mask, basic skincare routine, use of sunscreen, use of esthetician care, skincare concerns, skincare stress, and kind of cosmetic brand used, while there was a significant difference in the practical properties only by skin stress.

세안화장품 사용에 따른 20대 여대생의 피부상태 연구

임선형,곽선순,김애경 한국피부과학연구원 2010 대한피부미용학회지 Vol.8 No.4

This study analyzed skin conditions by cleanser types and investigated subjective satisfaction following each test in order to see the effects of cleansers applied at the cleansing stage deciding effectiveness of general skin care on the skin of female university students in their twenties. 28 female university students in Seoul, who were now found to have no skin disease, no acne, no edematous skin disease, and no dermatitis by inquiries and ocular inspections, participated in this study; for this purpose of measuring skin conditions accurately by cleanser types, they were asked to use the same blemish balm cream by Company A and compressed foundation by Company P as before the test. As for methodology, all of the 28 participants were asked to use the selected four types, such as cleansing lotion, cleansing oil, cleansing water, and cleansing gel, which needed no double cleansing, and then moisture, oil, pH, and the moisture evaporation rate in the skin were examined by using MPA5 580. Moisture and oil was highest with cleansing oil and lowest with cleaning water. pH was highest with cleansing gel and lowest with cleansing water. The moisture evaporation rate was highest with cleansing lotion and lowest with cleansing water, which was not significant. The results of this study demonstrate that cleansing oil containing much oil is good for dry and aged skin, while cleansing water and cleansing gel containing little oil are good for oily and acne-prone skin. This is identical with the purpose of this study that selection of a cleanser must depend on skin types. 본 연구는 피부미용에서 전체 피부관리의 효과를 좌우하는 세안단계에 사용되어지는 클렌저를 20대 여대생에게 사용한 후 피부상태를 살펴보기 위하여 클렌저제형별에 따른 피부상태를 분석하였으며 매 실험 후 주관적 만족도를 조사하였다. 연구 대상자는 서울 지역 소재 4년제 대학교에 재학 중인 여대생 중 현재 피부질환이 있거나 여드름이 있는 경우, 그리고 수종 피부질환이나 피부염증을 갖고 있는 사람은 문진과 시진으로 제외한 지원자 28명이 연구에 참여 하였으며, 클렌저제형에 따른 피부상태를 정확하게 측정하기 위해 실험 전 동일한 A사 Blemish Balm크림과 P사 압축 파운데이션을 사용하게 하였다. 본 논문은 O/W 형태의 4가지 세안제인 클렌징로션, 클렌징오일, 클렌징워터, 클렌징젤을 선정하여 피부의 수분, 유분, pH, 수분증발량을 MPA5 580을 이용하여 측정하였으며, 실험 전 자신에게 맞는 세안제 선택과 실험 후 만족하는 세안제를 조사한 결과 다음과 같은 결과를 얻었다. 수분과 유분은 ‘클렌징오일’ 가장 높게 나타났으며, ‘클렌징워터’ 가장 낮게 나타났다. pH는 ‘클렌징젤’이 가장 높게 나타났으며, ‘클렌징워터’가 가장 낮게 나타났다. 수분증발량은 ‘클렌징로션’이 가장 높게 나타났으며, ‘클렌징워터’가 가장 낮게 나타났으나 이 차이는 유의미하지 않는 것으로 나타났다. 연구결과에서 유분이 높은 ‘클렌징오일’은 ‘건성피부’, ‘노화피부’에 효과가 있으며, 유분이 적은 ‘클렌징워터’, ‘클렌징젤’은 ‘지성피부’, ‘여드름피부’에 효과가 있는 것으로 나타났다. 또한 20대 여대생은 세안용화장품으로 ‘클렌징로션’을 자신에게 맞는 세안제로 생각하였지만 실험 후 만족도 결과 ‘클렌징젤’을 가장 선호 하는 것으로 나타났다. 본 연구결과로 효과적인 피부관리를 위해서는 올바른 세안습관과 피부유형에 따른 효과 있는 세안화장품 선택을 해야 한다고 사료되었다.