• KSBB Journal
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• 제10권5호
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• pp.499-509
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• 1995

PU.1은 6개의 특이적인 purine-rich 염기서열 (5' -GAGGAA-3 )로 구성된 PU box에 결합하는 transcription activator이다. 이 PUol은 macro phage와 B-cell에서만 발현되어 이들 세포를 활성 화시키므로, 포유통물의 연역계를 연구하는 데 중요 한 위치를 차지하고 있다. Full length PUol cDNA 는 open reading frame 816개의 DNA 염기로 구성 되어 있으므로, 아미노산 2727~의 합성을 지령한다. PUol의 활성화는 이를 구성하고 있는 polypeptide 중 세린 잔기가 인산화되어 전사인자로서 작용한다 고 추측된다. PU.1은 22개의 세린을 함유하고 있으 며, 정확한 인산화 위치 빛 수량은 알 수 없으나, casein kinase II 에 의하여 인산화된다고 추측되는 제41,45,132'133,148번째 아미노산 세린들이 제1 차 target sites이다. 본 연구에서는 이상의 제41, 45, 132,133, 148번 아미노산 세린 codon(AGC, AGC, AGC.TCA, TCT)이 알라닌 codon(GCC, GCC, GCC.GCA, G GCT)으로 치환된 4가지의 점돌연변이체 클론 (pKKS41A, pRKS45A, pMKS132$.$133A, pMKS­1 148A)을 다음과 같이 제조하였다. Wild type PUol cDNA(template)를 해당되는 mutant DNA primers로 증폭(PCRjSOE)하여 mutant cDNA 단편을 얻었다. 이를 Hind III와 Xba I 으로 절단된 pBlu­e escript KS +에 접합시킨 후, 대장균(E. coli XLI ~ Blue)에 형질전환시켰다. 이 점돌연변이체들은 인산화 부위 및 수량은 물론 PU.1의 구조 및 기능 (Structure and Function) 연구에 기여할 것이다.

• 한국육종학회지
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• 제49권3호
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• pp.180-189
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• 2017

현재까지 국내에서는 생명공학작물이 상업적인 재배가 되고 있지 않으나 생명공학작물의 환경 방출을 위해서는 환경위해성 평가가 필수적 수행되어야 한다. 본 연구에서는 해충저항성 Bt 벼(Agb0101)로부터 모품종인 낙동벼와 잡초성벼인 R55 및 인디카벼인 IR36 로의 화분 매개에 의한 유전자 이동성을 평가하였다. 낙동벼로부터 729,917 립의 종자를 얻었으며, 잡초성벼(R55)로부터는 230,635 립의 종자와 인디카벼(IR36)에서는 596,318 립의 종자를 수확하였다. 교잡개체는 3회의 제초제 살포를 수행하여 제초제 저항성 개체 선별과 Cry1Ac1 immunostrip 검정으로 확인하였으며, 해충저항성 Bt 벼(Agb0101)에 특이적인 프라이머를 이용한 분자생물학적 방법을 통해 유전자 이동성 여부를 최종적으로 검증하였다. 총 파종된 종자수에 대한 교잡율은 낙동벼에서는 0.0027%, R55는 0.0017%, IR36은 0.0005%로 나타났으며, 모든 교잡개체들은 해충저항성 Bt 벼(Agb0101)에 근접한 1.2m 내에서 발견되었다. 화분 매개에 의한 해충저항성 Bt 벼(Agb0101)의 유전자 이동 특성은 기존에 연구된 결과들과 유사한 경향으로 보였으며, 벼의 개화기간 중 온도와 강우량 등 기상 조건이 화분에 의한 교잡을 결정하는데 중요한 요인으로 작용하였다. 이에 재배 지역의 기상 환경과 개화시기 중복 여부 등을 유전자변형 벼에 의한 일반 재배품종 및 야생(잡초성)벼로의 유전자 이동에 따른 저감 기술 개발과 안전관리 기준 작성에서 주요 영향 인자들로 고려되어야 할 것이다.

• ALGAE
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• 제34권2호
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• pp.111-126
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• 2019

The phototrophic dinoflagellate Biecheleriopsis adriatica is a small suessioid species characterized by a fragile thin wall. Although the morphology of this dinoflagellate is well established, there is currently little information available on its distribution and the environmental factors that influence this distribution. Thus, to investigate the spatial and seasonal distributions of the vegetative cells of B. adriatica in Korean waters, surface water samples were collected on a seasonal basis from 28 stations in the East, West, and South Sea of Korea and Jeju Island from April 2015 to October 2018, and abundances of the vegetative cells of B. adriatica were quantified using quantitative real-time polymerase chain reactions, for which we developed the species-specific primer and probe set. Simultaneously, major environmental parameters, including temperature, salinity, nutrient concentrations, and dissolved oxygen concentrations were measured. The vegetative cells of B. adriatica were detected at 20 of the 28 sampling stations: 19 stations in summer and 6 in autumn, although from no stations in either spring or winter. The ranges of water temperature and salinity at sites where this species was detected were $17.7-26.4^{\circ}C$ and 9.9-34.3, respectively, whereas those of nitrate and phosphate concentrations were not detectable-96.2 and $0.18-2.66{\mu}M$, respectively. Thus, the sites at which this species is found are characterized by a narrow range of temperature, but wide ranges of salinity and concentrations of nitrate and phosphate. The highest abundance of the vegetative cells of B. adriatica was $41.7cells\;mL^{-1}$, which was recorded in Jinhae Bay in July 2018. In Jinhae Bay, the abundance of vegetative cells was significantly positively correlated with the concentration of nitrate, but was negatively correlated with salinity. On the basis of these findings, it appears that the abundance of B. adriatica vegetative cells shows strong seasonality, and in Jinhae Bay, could be affected by the concentrations of nitrate.

• ALGAE
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• 제34권1호
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• pp.7-21
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• 2019

The genus Heterocapsa is one of the major dinoflagellate groups, with some of its species having worldwide distributions. However, prior to the present study, the phototrophic species Heterocapsa minima has been reported only from the northeast Atlantic Ocean. Recently, H. minima was found in the Korean waters, and a clonal culture was established. This culture was used to examine the morphology of the Korean strain H. minima HMMJ1604 through light and scanning electron microscopy, as well as for its genetic characterization. Furthermore, to determine the nationwide distribution of H. minima in Korea, its abundance was quantified in the waters of 28 stations in all four seasons in 2016-2018 using the quantitative real-time polymerase chain reaction method. The overall morphology of H. minima HMMJ1604 was very similar to that of the Irish strain H. minima JK2. However, the Korean strain had five pores around the pore plate, whereas the Irish strain had six pores. When properly aligned, the sequences of the large subunit and internal transcribed spacer regions of the ribosomal DNA of the Korean strain were identical to those of the Irish strain. This species was detected in the waters of 26 out of 28 stations, but its abundance was greater than $1.0cells\;mL^{-1}$ at 8 stations. The highest abundance of H. minima was $44.4cells\;mL^{-1}$. Although this species was found in all seasons, its abundance was greater than $1.0cells\;mL^{-1}$ when the water temperature and salinity were $10.9-25.0^{\circ}C$ and 17.5-34.1, respectively. To the best knowledge, the present study reported for the first time that H. minima lives in the Pacific Ocean and is widely distributed in the Korean waters.

• 식물병연구
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• 제26권4호
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• pp.283-288
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• 2020

2017년 9월 충남 예산지역의 아욱 잎에서 엽맥퇴록 및 황화 등 바이러스 증상 관찰되었다. 증상이 있는 아욱 5주에서 핵산을 추출하여 아욱엽맥투명바이러스(Malva vein clearing virus, MVCV)를 포함한 4종 바이러스 특이 프라이머를 이용하여 reverse transcription polymerase chain reaction 수행하였다. MVCV 특이 프라이머로 증폭된 total RNA 5점에서 600 bp의 분자크기를 갖는 밴드가 확인되었다. MVCV 확인하기 위하여 여기서 얻어진 PCR 산물 3개를 정제 후 direct sequencing으로 염기서열을 결정하였다. BLAST 검색 결과, 멕시코의 토마토에서 분리된 MVCV 분리주와 99%로 가장 높은 상동성을 보였다. 명아주속 식물을 이용하여 단일국부병반을 3회 분리 후 Cm1-5으로 명명하였다. Cm1, Cm3, Cm5 분리주를 23종의 지표식물에 즙액 접종하였다. Cm3 분리주는 접시꽃에 퇴록반점 및 모자이크 증상을 나타내었다. 국내 아욱 3 분리주를 포함한 6개 다른 국가 및 식물 종에서 분리된 19 MVCV 분리주들에 대한 외피 단백질 유전자의 계통학적 유연관계분석 결과, 지리적 기원 또는 병원성과는 관련되지 않았다. 본 연구는 국내 아욱에서 MVCV의 자연발생 및 3 분리주의 특성에 대한 첫 보고이다.

• ALGAE
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• 제36권1호
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• pp.37-50
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• 2021

Heterotrophic dinoflagellates Gyrodinium spp. are one of the major grazers of phytoplankton in many coastal waters. Gyrodinium dominans, G. jinhaense, and G. moestrupii have similar morphologies but different edible prey species. To explore the variations in the ecological niches of these three species, we investigated their spatial-temporal distributions in Korean waters. Because of the high similarity in morphology among these three Gyrodinium species, we used real-time polymerase chain reactions to quantify their abundance in water samples that were seasonally collected from 28 stations along the Korean Peninsula from April 2015 to October 2018. Cells of G. dominans were found at all sampling stations, G. jinhaense at 26 stations, and G. moestrupii at 22 stations, indicating that all three species were widely distributed in Korea. Furthermore, all three species displayed strong seasonal distributions. The largest numbers of the stations where G. dominans and G. jinhaense cells were present were found during the summer (26 and 23 stations, respectively), but that for G. moestrupii was found in the autumn (15 stations). The abundance of G. dominans was positively correlated with that of G. jinhaense, but not with that of G. moestrupii. The highest abundances of G. dominans (202.5 cells mL-1) and G. jinhaense (20.2 cells mL-1) were much greater than that of G. moestrupii (1.2 cells mL-1). The highest abundances of G. dominans and G. jinhaense were found in July, whereas that of G. moestrupii was found in March. The abundances of G. dominans and G. jinhaense, but not G. moestrupii, were positively correlated with water temperature. Therefore, the spatial-temporal distributions of G. dominans and G. jinhaense were closer than those of G. moestrupii and G. dominans or G. jinhaense. This differs from results based on the relative differences in ribosomal DNA sequences and the types of edible prey reported in the literature. Thus, the variations in spatial-temporal distributions and prey species of these three Gyrodinium species suggest that they may have different ecological niches in Korean coastal waters.

• 대한본초학회지
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• 제27권6호
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• pp.37-42
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• 2012

Objectives : Angelica Gigantis Radix is prescribed as the root of different Angelica species on the pharmacopoeia in Korea, Japan and China. Chemical components and their biological activities were also different according to their species. A study for the development of simple method to compare Angelica roots was needed. In order to classify them, the methods such as DNA sequencing analysis and taste sensor were applied to three Angelica species like Angelica gigas, Angelica acutiloba and Angelica sinensis. Methods : PCR amplification of intergenic transcribed spacer (ITS) region was performed using ITS1 and ITS4 primer from nine Angelica roots, and then nucleotide sequence was determined. Taste pattern of samples were measured using the taste-sensing system SA402B equipped with a sensing unit, which consists of artificial lipid membrane sensor probes of anionic bitterness, astringency, saltiness, umami, and cationic bitterness (C00, AE1, CT0, AAE, and AN0, respectively). Results : As a result of comparing the similarity of the ITS region sequences, A. sinensis was discriminated from the others (A. gigas and A. acutiloba). Equally this genetic result, A. gigas and A. acutiloba showed similar taste pattern as compared to A. sinensis. Sourness, bitterness, aftertaste of bitterness, astringency, and aftertaste of astringency of A. sinensis were significantly high as compared with A. gigas and A. acutiloba. In contrast, richness was significantly low. Conclusions : These taste pattern can be used as a way of comparison of Angelica species and this technic could be applied to establish a taste pattern marker for standardization of herbs in various purposes.

• 생명과학회지
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• 제33권8호
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• pp.623-631
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• 2023

본 연구는 차세대 염기서열 분석법(NGS)을 사용하여 벤자리의 microsatellite 마커를 개발하고, 개발한 마커를 사용하여 벤자리 집단의 유전학적 특성을 분석하기 위해 수행되었다. 차세대 염기서열 분석 장비인 Illumina Hiseq X ten을 사용하여 총 402,244,934개의 read들을 얻어 assembly를 실행한 결과, 전체의 약 0.33%에 해당하는 1,320,995개의 read들이 assembly 되었으며, 이 read들의 총 길이는 705,613,658 bp로 확인되었다. 크기가 640 bp 이상이 되는 contig는 952,326개로 나타났으며, 이 중 microsatellite 영역을 포함하는 contig를 151개(0.016%)로 1차 선별하고, PCR 증폭 여부를 통해 microsatellite 후보 34개를 2차로 선별하였다. 그 중 벤자리 집단의 마커로서 유용한 15개의 microsatellite 마커를 최종 선택하였다. 새로 개발한 15개의 microsatellite 마커를 사용하여 벤자리 집단을 대상으로 분석한 결과, 관찰된 유효 대립유전자 수(N_A)는 평균 12(6~25)로 나타났다. 평균 관측치 이형접합도(H_O)와 평균 기대치 이형접합도(H_E)는 각각 0.750(0.530-0.873)와 0.793 (0.647-0.895)으로 나타냈으며, 이는 해산어의 평균 값인 0.79와 유사한 수치를 나타내었다. 따라서 본 연구에서 개발한 15개의 microsatellite 마커는 벤자리 집단의 유전학적 특성 분석에 유용할 것으로 사료된다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.46-46
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• 2003

Interleukin-10 (IL-10) is a homodimeric protein with a wide spectrum of anti-inflammatory and immune activities. It inhibits cytokine production and expression of immune surface molecules in various cell types. The transgenic mice carrying the human IL-10 gene in conjunction with the bovine $\beta$-casein promoter produced the human IL-10 in milk during lactation. Transgenic mice were generated using a standard method as described previously. To screen transgenic mice, PCR was carried out using chromosomal DNA extracted from tail or toe tissues with a primer set. In this study, stability of germ line transmission and expression of IL-10 gene integrated into host chromosome were monitored up to generation F15 of a transgenic line. When female mouse of generation F9 was crossbred with normal male, generation F9 to F15 mice showed similar transmission rates (66.0$\pm$20.13%, 61.5$\pm$16.66%, 41.1$\pm$8.40%, 40.7$\pm$20.34%, 61.3$\pm$10.75%, 49.2$\pm$18.82%, and 43.8$\pm$25.91%, respectively), implying that the IL-10 gene can be transmitted stably up to long term generation in the transgenic mice. For ELISA analysis, IL-10 expression levels were determined with an hIL-10 ELISA and a mIL-10 ELISA kit in accordance with the supplier's protocol. Expression levels of human IL-10 from milk of generation F9 to F13 mice were 3.6$\pm$1.20 mg/ml, 4.2$\pm$0.93 mg/ml, 5.7$\pm$1.46 mg/ml, 6.3$\pm$3.46 mg/ml, and 6.8$\pm$4.52 mg/ml, respectively. These expression levels are higher than in generation F1 (1.6 mg/ml) mice. We concluded that transgenic mice faithfully passed the transgene on their progeny and successively secreted target proteins into their milk through several generations, although there was a little fluctuation in the transmission frequency and expression level between the generations.

• 대한본초학회지
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• 제24권3호
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• pp.59-68
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• 2009

Objectives : Bupleuri Radix (Siho) is prescribed as the root of different Bupleurum species on the pharmarcopoeia in Korea and China. Moreover, other species and varieties of the genus Bupleurum have been also distributed on the herbal market as Bupleuri Radix. However, due to the morphological similarity and frequent occurrence of intermediate forms, the correct identification of this radix is very difficult. To develop a reliable method for correct identification and improving the quality standards of official Bupleuri Radix, we analyzed sequences of the ribosomal RNA gene and internal transcribed spacer (rDNA-ITS) region. Methods : PCR amplification of rDNA-ITS region was performed using ITS1 and ITS4 primer from 6 Bupleurum species and 1 variety, B. falcatum L. (Siho), an improved breed of B. falcatum L. (Samdo-Siho), B. chinense DC. (Buk-Siho), B. scorzonerifolium Willd. (Nam-Siho), B. longiadiatum Turcz. (Gae-Siho), B. euphorbiodes Nakai (Deungdae-Siho) and B. latissimum Nakai (Seom-Siho), and nucleotide sequence was determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW using entire rDNA-ITS sequence of three samples per species. Results : In comparative analysis of the rDNA-ITS sequences, we found specific nucleotides to distinguish Korean (B. falcatum L. and its variety) and Chinese official species (B. chinense DC. and B. scorzonerifolium Willd.) from others at positions 411 and 447, and positions 89, 101, 415 and 599, respectively. Futhermore, we also found nucleotide indels (insertion and/or deletion) and substitutions to identify each of different Bupleurum species, 2 positions for B. falcatum L. and its variety, 6 positions for B. chinense DC., 49 positions for B. scorzonerifolium Willd., 8 positions for B. euphorbioides Nakai, 7 positions for B. longiradiatum Nakai and 9 positions for B. latissimum Nakai. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origins of Bupleuri Radix. Moreover, we confirmed the phylogenetic relationship of B. latissimum Nakai, a Korean endemic speices, among Bupleurum species based on the rDNA-ITS sequence. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterant Bupleurum species.