• 한국약용작물학회지
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• 제12권1호
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• pp.53-59
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• 2004

국내 한약재 시장에서 유통되고 있는 백출 건재약재의 기원을 판별하고자 해부형태적 특징과 종 특이적인 SCAR 마커 (SAjR2, SAmR1)를 적용한 실험 결과는 다음과 같다. 해부형태적 특징으로 A. japonica에서는 주피에 코르크층이 매우 발달하여 있었고, 코르크층 내부 또는 아래에 석세포대가 연속적으로 발달하고 있었다. 유실은 A. japonica에서 뿌리횡단면에 전체적으로 다수 분포하여 잘 발달하고 있었으며 피충부분과 잘 발달된 방사조직 내에서도 목섬유다발이 다수 관찰됐다. 또한 유세포 내의 침상 결정물도 A. macrocephala보다 많이 함유하고 있었다. 이러한 특징들을 종합한 결과, 국내 유통 국산 및 수입 백출은 모두 A. japonica의 해부형태적 특징을 따르고 있어 A. japonica로 동정되었다. 한편 RAPD clone으로부터 개발된 A. japonica 특이적인 SAjR2를 이용하여 PCR 한 결과 모든 샘플에서 600bp 크기의 한국백출 특이적인 밴드가 형성되었으며, A. macrocephala 특이적인 SAmR1을 이 용한 결과에서는 작물시험장에서 재배중인 A. macrocephala에서만 1200 bp크기의 특이적인 밴드가 형성되어, 시중에서 유통되고 있는 한국백출과 중국백출은 모두 A. japonica로 동정되었다. 결론적으로 해부형태적 특성과 SCAR 마커는 A. japonica와 A. macrocephala 기원식물의 동정과 유통 건재 약재를 판별하기에 유용한 방법이었다.

• Reproductive and Developmental Biology
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• 제28권4호
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• pp.247-252
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• 2004

Human embryonic stem (ES) cells are derived from the inner cell mass of the preimplantation embryo. Human ES cells have the capacity to differentiate into various types of cells in the body. Human ES cells are indefinite source of cells for cell therapy in various degenerative disorders including neuronal disorders. Directed differentiation of human ES cells is a prerequisite for their clinical application. The objective of this study is to develop the culture condition for the derivation of neural precursor cells from human ES cells. Neural precursor cells were derived from human ES cells in a stepwise culture condition. Neural precursor cells in the form of neural rosette structures developed into neurospheres when cultured in suspension. Suspension culture of neurospheres has been maintained over 4 months. Expressions of nestin, soxl, sox2, pax3 and pax6 transcripts were upregulated during differentiation into neural precursor cells by RT-PCR analysis. In contrast, expression of oct4 was dramatically downregulated in neural precursor cells. Immunocytochemical analyses of neural precursor cells demonstrated expression of nestin and SOX1. When induced to differentiate on an adhesive substrate, neuro-spheres were able to differentiate into three lineages of neural systems, including neurons, astrocytes and oligo-dendrocytes. Transcripts of sox1 and pax6 were downregulated during differentiation of neural precursor cells into neurons. In contrast, expression of map2ab was elevated in the differentiated cells, relative to those in neural precursor cells. Neurons derived from neural precursor cells expressed NCAM, Tuj1, MAP2ab, NeuN and NF200 in immunocytochemical analyses. Presence of astrocytes was confirmed by expression of GFAP immuno-cytochemically. Oligodendrocytes were also observed by positive immuno-reactivities against oligodendrocyte marker O1. Results of this study demonstrate that a stepwise culture condition is developed for the derivation of neural precursor cells from human ES cells.

• Reproductive and Developmental Biology
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• 제36권1호
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• pp.33-37
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• 2012

Differential capacity of the parthenogenetic embryonic stem cells (PESCs) is still under controversy and the mechanisms of its neural induction are yet poorly understood. Here we demonstrated neural lineage induction of PESCs by addition of insulin-like growth factor-2 (Igf2), which is an important factor for embryo organ development and a paternally expressed imprinting gene. Murine PESCs were aggregated to embryoid bodies (EBs) by suspension culture under the leukemia inhibitory factor-free condition for 4 days. To test the effect of exogenous Igf2, 30 ng/ml of Igf2 was supplemented to EBs induction medium. Then neural induction was carried out with serum-free medium containing insulin, transferrin, selenium, and fibronectin complex (ITSFn) for 12 days. Normal murine embryonic stem cells derived from fertilized embryos (ESCs) were used as the control group. Neural potential of differentiated PESCs and ESCs were analyzed by immunofluorescent labeling and real-time PCR assay (Nestin, neural progenitor marker; Tuj1, neuronal cell marker; GFAP, glial cell marker). The differentiated cells from both ESC and PESC showed heterogeneous population of Nestin, Tuj1, and GFAP positive cells. In terms of the level of gene expression, PESC showed 4 times higher level of GFAP expression than ESCs. After exposure to Igf2, the expression level of GFAP decreased both in derivatives of PESCs and ESCs. Interestingly, the expression level of $Tuj1$ increased only in ESCs, not in PESCs. The results show that IGF2 is a positive effector for suppressing over-expressed glial differentiation during neural induction of PESCs and for promoting neuronal differentiation of ESCs, while exogenous Igf2 could not accelerate the neuronal differentiation of PESCs. Although exogenous Igf2 promotes neuronal differentiation of normal ESCs, expression of endogenous $Igf2$ may be critical for initiating neuronal differentiation of pluripotent stem cells. The findings may contribute to understanding of the relationship between imprinting mechanism and neural differentiation and its application to neural tissue repair in the future.

• 생명과학회지
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• 제16권6호
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• pp.1001-1005
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• 2006

SSR markers를 이용하여 벼의 품종간 유전적 유연관계 분석과 품종식별 방법에 대한 연구를 수행하여 얻어진 결과를 요약하면 다음과 같다. SSR primer 50개와 벼 보급종 21품종을 PCR 반응시킨 결과 다형성을 뚜렷하게 나타내는 primer는 23개였으며, 각 marker에 의해 발생된 대립유전자의 수는 $2{\sim}9$까지 검출되었고, 평균값은 3.00개로 나타났다. 유전적 다형성 정도를 나타내어 주는 SSR marker의 PIC 값은 최소 0.091에서부터 최대 0.839까지 다양하게 분석되었다. SSR marker를 이용하여 분석된 벼 21품종에 대한 전체 유전적 유사도는 $0.59{\sim}0.92$의 범위에 속하였고 유사도 지수 0.65를 기준으로 할 때 4개의 그룹으로 구분되었다. SSR marker중에서 RM206, RM225, RM418, RM478은 marker genotype에 의해 21 품종에 대해 각각 고유한 밴드 특성을 나타내어 품종판별이 가능한 것으로 나타났다. 금후 이 연구결과는 벼 보급종의 품종식별을 위해 효과적으로 이용될 수 있는 것으로 나타났다.

• 생명과학회지
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• 제30권7호
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• pp.614-624
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• 2020

최근 next-generation sequencing technology의 발달로 유전체 분석 사례는 증가하고 있으나, 단감에 있어 적용 가능한 Single Nucleotide Polymorphism (SNP) 마커 및 적용 결과는 거의 없는 실정이다. 이에 우리나라 고유 떫은감 5품종으로부터 개발된 SNP primer 들을 단감 품종에 적용하여 사용 가능성을 검증하고자 수행하였다. Jung 등에 의해 개발된 19개 SNP primer들의 PCR 조건을 확인 한 후 본 실험의 전기영동 방식으로는 분석이 매우 어려웠던 8개의 primer를 제외한 11개의 SNP primer들을 최종 선발하였다. 1, 2차 검증을 통해 최종 선발된 11개의 SNP primer 들을 76품종 및 계통(불완전단감 20, 완전단감 30, 완전떫은감 20, 불완전떫은감 6)에 적용한 결과 38품종 및 계통(불완전단감 8, 완전단감 18, 완전떫은감 9, 불완전떫은감 3품종)은 각 품종 및 계통 간 구분을 할 수가 없었다. 그러나 최종 선발된 11개의 SNP primer 들을 신품종에 적용한 결과만를 보면 '감누리', '단누리', '홍추'와 '자미시', '미감조생'을 동시에 구분할 수 있어 단감 신품종 판별을 위한 특이적 마커로 사용될 수 있을 것으로 판단된다.

• 한국가금학회지
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• 제40권3호
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• pp.223-234
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• 2013

Poultry industry is abounding day by day as it engrosses less cost of investment per bird as compared to large animals. Poultry have the most copious genomic tool box amongst domestic animals for the detection of quantitative trait loci (QTL) and marker assisted selection (MAS). Use of multiple markers and least square techniques for mapping of QTL affecting quality and production traits in poultry is in vogue. Examples of genetic tests that are available to or used in industry programs are documented and classified into causative mutations (direct markers), linked markers in population-wide linkage disequilibrium (LD) with the QTL (LD markers), and linked markers in population wide equilibrium with the QTL (LE markers). Development of genome-wide SNP assays, role of 42 K, 60 K (Illumina) and 600 K (Affymetrix$^{(R)}$ Axim$^{(R)}$) SNP chip with next generation sequencing for identification of single nucleotide polymorphism (SNP) has been documented. Hybridization based, PCR based, DNA chip and sequencing based are the major segments of DNA markers which help in conducting of MAS in poultry. Economic index-marker assisted selection (EI-MAS) provides platform for simultaneous selection for production traits while giving due weightage to their marginal economic values by calculating predicted breeding value, using information on DNA markers which are normally associated with relevant QTL. Understanding of linkage equilibrium, linkage dis-equilibrium, relation between the markers and gene of interest are quite important for success of MAS. This kind of selection is the most useful tool in enhancing disease resistance by identifying candidate genes to improve the immune response. The application of marker assisted selection in selection procedures would help in improvement of economic traits in poultry.

• 대한본초학회지
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• 제35권4호
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• pp.51-60
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• 2020

Objective : The objective of this study was to demonstrate the effect of Persicae Semen (PS) in DNCB-induced atopic dermatitis mouse and HaCaT cell. Methods : The BALB/c mice were divided into four groups. To develop atopic dermatitis, 200 ㎕ of 1 and 0.5% DNCB solution was put on the back of mice in the Control group, the PS-Low group and the PS-High group once a day. After application of DNCB, 200 ㎕ of the PS extract was also treated. The Normal group was given PBS. The mice dorsal skin was stained with Masson's trichrome, H&E, and toluidine blue to evaluate the thickness of the epidermis and dermis, infiltration of eosinophils and mast cells respectively. ELISA was applied to measure the serum level of IgE and IL-6. Toxicity of PS was measured by MTS assay in HaCaT cell. To investigate the effects of PS on HaCaT cells, cells were pre-treated with PS for 1h, and then stimulated with TNF-α and IFN-γ. After 24 hours, the expression of TARC was analyzed using RT-PCR. Results : PS not only significantly diminished the thickness of the epidermis and dermis, but also reduced the infiltration of eosinophil and mast cell in skin lesion. PS also reduced the serum IgE and IL-6 level which plated important roles in the atopic dermatitis. The expression of TARC was decreased significantly in TNF-α/IFN-γ stimulated HaCaT cell. Conclusion : These results suggest that PS may be effective in alleviating the atopic dermatitis induced by DNCB and inflammation by TNF-α/IFN-γ.

• 구강회복응용과학지
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• 제36권3호
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• pp.145-157
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• 2020

목적: 본 연구에서는 구강연조직재생에서 인간치은섬유아세포에 hydrophobically modified glycol chitosan (HGC) 기반 나노방출제어시스템을 적용 시 PI3K-AKT 신호전달의 세포생존 연관 유전자를 통해 trichloroacetic acid (TCA)와 epidermal growth factor (EGF)의 영향을 확인하고자 하였다. 연구 재료 및 방법: TCA와 EGF를 방출하는 나노방출제어시스템을 제작하였다. 실험군은 3가지 군으로 나누었다; 대조군(CON), TCA-담지형 나노방출제어시스템 투여군(EXP1), TCA-와 EGF- 담지형 나노방출제어시스템 투여군(EXP2). 인간치은섬유아세포 배양 후 PI3K-AKT 신호전달에 연관된 총 29개 유전자를 실시간 중합효소연쇄반응으로 분석했다. 일요인 분산분석 및 다중회귀분석이 사용되었다. 결과: 세포생존 관련 유전자들은 EXP1과 EXP2에서 상향조절되었다. 다중회귀분석에서는 ITGB1이 AKT1의 발현에 가장 영향력 있는 요소로 결정되었다. 결론: HGC기반 나노방출제어시스템을 통한 TCA와 EGF의 적용은 세포생존에 관한 신호전달을 상향 조절시킬 수 있다.

• 한국수정란이식학회지
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• 제29권3호
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• pp.297-303
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• 2014

The objective of this study was to determine the breed differences in the 3'-untranslated region (UTR) of MC1R mRNA, which may be used to distinguish Korean brindle cattle (Chikso) from other breeds. We investigated the relationship between the variation of 3'-UTR of the MC1R mRNA and coat color among different breeds and the Korean brindle cattle with different coat colors. MC1R mRNA expression levels were determined in accordance with the coat color and hair colors of the tail. Total cellular RNA was extracted from the hair follicles of the tails in Hanwoo, Korean brindle cattle, Holstein and $Hanwoo{\times}Holstein$ crossbred cattle. After cDNA synthesis, PCR was performed. Sequences of the 3'-UTR of MC1R mRNA were analyzed. The 3'-UTR of the MC1R mRNA from different breeds of cattle did not show any variations. There were no variations in the 3'-UTR of the MC1R mRNA in Korean brindle cattle with different coat colors. The levels of MC1R mRNA expression in hair follicles of the tail varied substantially among the Korean brindle cattle with different coat colors, except yellow coat color. Correlation between the MC1R mRNA expression in the hair follicles of the tail and coat color may be present in the Korean brindle cattle, but not between the variations of 3'-UTR of MC1R mRNA and coat color. Further studies to determine the regulation of MC1R mRNA expression from the hair follicles of different coat colors will be beneficial in clarifying the role of MC1R in the coat colors of the Korean brindle cattle.

• 한국수정란이식학회지
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• 제32권3호
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• pp.73-79
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• 2017

Transplantation is considered to be a very useful approach to improve human welfare and to prolong life-span. Heterologous organ transplantation using pig organs which are similar to human beings and easy to make mass-production has known as one of the alternatives. To ensure potential usage of the pig organ for transplantation application, it is essentially required to generate transgenic pig modifying immuno-related genes. Previously, we reported production of heterozygous ${\alpha}1,3$-galactosyltransferase (GalT) knock-out and human membrane cofactor protein (MCP) expressing pig ($GalT^{-MCP/+}$), which is enforced for suppression of hyperacute and acute immunological rejection. In this study, we reported generation of homozygous pig ($GalT^{-MCP/-MCP}$) by crossbreeding $GalT^{-MCP/+}$ pigs. Two female founders gave birth to six of $GalT^{-MCP/-MCP}$, and seven $GalT^{-MCP/+}$ pigs. We performed quantitative real-time PCR, western blot, and flow cytometry analyses to confirm GalT and MCP expression. We showed that fibroblasts of the $GalT^{-MCP/-MCP}$ pig do not express GalT and its product Gal antigen, while efficiently express MCP. We also showed no expression of GalT, otherwise expression of MCP at heart, kidney, liver and pancreas of transgenic pig. Taken together, we suggest that the $GalT^{-MCP/-MCP}$ pig is a useful candidate to apply xenotransplantation study.