瘀血에 應用되는 當歸飮의 消炎, 解熱, 鎭痛作用에 미치는 影響

閔炳煥,蔡炳允 慶熙大學校韓醫科大學韓醫學硏究所 1983 慶熙韓醫大論文集 Vol.6 No.-

Stock(Matthiola incana R. Br.)으로부터 색소유전자의 분리 및 분석

민병환,김석원,오승철,유장렬 한국식물생명공학회 1998 식물생명공학회지 Vol.25 No.5

색소유전자의 전이를 통하여 새로운 색소발현체계를 가진 품종을 육종하기 위한 기초연구로 stock (Matthiola incana R. Br.)의 꽃봉오리로부터 cDNA-library를 합성하였고 screening을 통하여 anthocyanin 합성경로의 중요효소의 하나인 DFR (dihydroflavonol 4-reductase) 유전자를 분리하였다. 염기서열분석을 수행하여 분리유전자의 크기가 1450bp 이며 이중 coding region은 1029 bp 임을 확인하였다. 이미 밝혀진 다른 식물체의 DFR 유전자와 서로 염기서열의 일치성을 비교해 본 결과 외자엽식물인 옥수수와 보리와는 각각 61%를 보였으며, 쌍자엽식물인 페튜니아, 금어초, 거베라, 과꽃 그리고 카네이션 등 과는 66%-67%의 일치성을 나타내었다. 아울러 염기서열의 G/C 함량분석을 통하여 쌍자엽식물의 G/C 함량은 외자엽식물의 그것에 비해 매우 낮은 수치를 나타내었다. 분리유전자의 발현을 확인하기 위하여 인위적으로 기내에서의 전사와 해석을 수행한 결과 42-44 kd 크기의 단백질을 확인하였다. Southern blot 분석의 결과 DFR 유전자는 stock의 genome에 다른 대부분의 식물체와 유사하게 한 개가 존재하며 야생종과 돌연변이종의 stock을 분리 DFR 유전자를 probe 로 Northern blot 분석을 수행하여 돌연변이종인 lineK17b가 DFR 돌연변이임을 확인하였다. In this paper we describe the cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme dihydroflavonol 4-reductase (DFR) in Matthiola incana R. Br. A heterologous cDNA probe from Zea mays was used to isolate full-size DFR cDNA clone from a corolla-specific cDNA library. Comparison of the coding region of this DFR cDNA sequence including the sequences of Zea mays, Anthirrinum majus, Petunia hybrida, Callistephus chinensis, Dianthus caryophyllus and Rosa hybrida reveals a identity higher than 61% at the nucleotide level. The DFR transcript is G/C rich in monocotyledonous plants show a strong codon bias preferring codons with a G or C in the third position. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRNA from wild type and mutant plants and by in vitro expression yielding an enzymatically active reductase. Genomic southern blot analysis showed the presence of one gene for DFR in Matthiola incana. Northern blot analysis of the DFR wild type and mutant lines showed that the lack of DFR activity in the stable acyanic mutant k17b is clearly by a transcriptional block of the DFR gene.

Cloning and Expression Study of Dihydroflavonol 4-Reductase from Summer Aster (Callistephus chinensis)

민병환 한국원예학회 2006 Horticulture, Environment, and Biotechnology Vol.47 No.4

Dihydroflavonol 4-reductase (DFR) is a key enzyme involved in anthocyanin biosynthesis in aster. DFR catalyses the reduction of dihydroflavonols to leucoanthocyanidins in the anthocyanin biosynthesis pathway. A heterologous cDNA probe from Petunia hybrida was used to isolate DFR-encoding cDNA clone from summer aster (Callistephus chinensis). Comparison of the coding region of this DFR cDNA sequence including the sequences of Zea mays, Anthirrinum majus, P. hybrida, Matthiola incana, Dianthus caryophyllus, and Rosa hybrida reveals a similarity higher than 61% at the nucleotide level. The DFR transcript is G/C rich in monocotyledonous plants show a strong codon bias preferring codons with a G or C in the third position. The function of this nucleotide sequence was verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by Northern blotting with mRNA from wild type and mutant plants, and by in vitro expression yielding an enzymatically active reductase. Genomic Southern blot analysis showed the presence of one gene for DFR in C. chinensis. Northern blot analysis of the DFR wild-type and mutant lines strongly suggested that the lack of DFR activity in the stable acyanic mutant 10h may be caused by a transcriptional block of the DFR gene.

pEA 9::Tn5-Mob에 의한 nif-plasmid pEA 9의 transfer 성질

민병환,이호자 한국미생물학회 1988 미생물학회지 Vol.26 No.3

Using a Tn5-Mob system, pEA9 was characterized as a self-transmissible plasmid carrying a kanamycin resistance marker. The self-transfer frequencies of pEA9 varied greatly depending on pH values. The transfer frequency was about $4\times 10^{-5}$ at pH 5, that was 10 times higher than one at pH 6.5. With a helper plasmid, transfer frequencies were increased about $10^{4}$ times than the frequencies obtained without it.

안개초(Gyposphila paniculata)로부터 Flavanone 3β-Hydroxylase 유전자의 분리 및 분석

민병환,Min, Byung-Whan 한국식물생명공학회 2006 식물생명공학회지 Vol.33 No.2

Flavanone 3$\beta$-hydroxylase (FHT)는 flavonoid 생합성 경로의 가장 중심부에 작용하는 효소로 flavanone으로부터 dihydroflavonol으로의 변환을 촉매하는 역할을 한다. 본 연구에서는 색소유전자의 전이를 통하여 새로운 색소발현체계를 가진 품종을 육종하기 위한 기초연구로 숙성안개초 (Gypsophila paniculata L.)의 꽃봉오리로부터 cDNA-library를 합성하였고 카네이션의 FHT 유전자를 probe로 사용하여 anthocyanin 합성경로의 중요 효소의 하나인 FHT 유전자를 분리하였다. 염기서열분석을 수행하여 분리유전자의 크기가 1471 bp 이며 이 중 coding region은 1047 bp 임을 확인하였다. 이미 밝혀진 다른 식물체의 FHT 유전자와 서로 염기서열의 일치성을 비교해 본 결과 아라비돕시스, 오렌지, 카네이션, 고구마, 스톡, 페튜니아, 감자 및 포도에서 각각 69% 이상을 나타내었다. 분리유전자의 발현을 확인하기 위하여 Northern blot분석 및 인위적으로 기내에서의 transcription과 translation을 수행하였고, 분리한 유전자의 효소활성을 측정해 본 결과 dihtydrokaempferol의 작은 peak을 확인하였다. Southern blot 분석의 결과 안개초의 FHT 유전자는 다른 대부분의 식물체와 유사하게 한 개가 존재함을 확인하였다 Flavanone 3$\beta$-hydroxylase (FHT) is an enzyme acting in the central part of the flavonoid biosynthesis pathway. FHT catalyses the hydroxylation of flavanone to dihydroflavonols in the anthocyanin pathway. In this paper we describe the cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme FHT in Gypsophila paniculata L. A heterologous cDHA probe from Dianthus cavophyllus was used to isolate FHT-encoding cDHA clones from Gypsophila paniculata L.. Inspection of the 1471 bp long sequence revealed an open reading frame 1047 bp, including a 190 bp 5' leader region and 288 bp 3' untranslated region. Comparison of the coding region of this FHT cDHA sequence including the sequences of Arabidopsis thaliana, Citrus sinensis, Dianthus caryophyllus, Ipomoea batatas, Matthiola incana, Nierembergia sp, Petunia hybrida, Solanum tuberosum, Vitis vinifera reveals a identity higher than 69% at the nucleotide level. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRHA from wild type and mutant plants, by in vitro expression yielding and enzymatically active hydroxylase, as indicated by the small dihydrokaempferol peak. Genomic southern blot analysis showed the presence of only one gene for FHT in Gypsophila paniculata.

안개초(Gypsophila paniculata)로부터 Flavanone 3β-Hydroxylase 유전자의 분리 및 분석

민병환 한국식물생명공학회 2006 JOURNAL OF PLANT BIOTECHNOLOGY Vol.33 No.2

Flavanone 3β-hydroxylase (FHT) is an enzyme acting in the central part of the flavonoid biosynthesis pathway. FHT catalyses the hydroxylation of flavanone to dihydroflavonols in the anthocyanin pathway. In this paper we describe the cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme FHT in Gypsophila paniculata L. A heterologous cDNA probe from Dianthus caryophyllus was used to isolate FHT-encoding cDNA clones from Gypsophila paniculata L.. Inspection of the 1471 bp long sequence revealed an open reading frame 1047 bp, including a 190 bp 5' leader region and 288 bp 3' untranslated region. Comparison of the coding region of this FHT cDNA sequence including the sequences of Arabidopsis thaliana, Citrus sinensis, Dianthus caryophyllus, Ipomoea batatas, Matthiola incana, Nierembergia sp, Petunia hybrida, Solanum tuberosum, Vitis vinifera reveals a identity higher than 69% at the nucleotide level. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRNA from wild type and mutant plants, by in vitro expression yielding and enzymatically active hydroxylase, as indicated by the small dihydrokaempferol peak. Genomic southern blot analysis showed the presence of only one gene for FHT in Gypsophila paniculata.

안개초(Gypsophila paniculata L.)로부터 dihydroflavonol 4-reductase 유전자의 분리 및 분석

민병환,정동춘 한국식물생명공학회 2010 식물생명공학회지 Vol.37 No.1

Dihydroflavonol 4-reductase (DFR) is a key enzyme of the flavonoid biosynthesis pathway which catalyses the NADPH-dependent reduction of 2R,3R-trans-dihydroflavonols to leucoanthocyanidins. In this study we describe cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme DFR in Gypsophila paniculata L. Inspection of the 1279 bp long sequence revealed an open reading frame 1063 bp, including a 36 bp 5' leader region and 181 bp 3' untranslated region. Comparison of the coding region of this DFR cDNA sequence including the sequences of Arabidopsis thaliana, Citrus sinensis, Dianthus caryophyllus,Ipomoea batatas, Matthiola incana, Nierembergia sp, Petunia hybrida, Solanum tuberosum, Vitis vinifera reveals an identity higher than 69% at the nucleotide level. The function of this nucleotide sequences was verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRNA from wild type and mutant plants, by in vitro expression yielding and enzymatically active reductase, as indicated by the small leucopelargonidin peak. Genomic southern blot analysis showed the presence of only one gene for DFR in Gypsophila paniculata. Dihydroflavonol 4-reductase(DFR)는 flavonoid 생합성 경로의 가장 중심부에 작용하는 효소로 2R,3R-trans-dihydroflavonols로부터leucoanthocyanidins 으로의 변환을 촉매한다. 본 연구에서는 색소유전자의 전이를 통하여 새로운색소발현체계를 가진 품종을 육종하기 위한 기초연구로안개초 (Gypsophila paniculata L.)의 꽃봉오리로부터 cDNAlibrary를합성하였고 카네이션의 DFR 유전자를 probe로사용하여 anthocyanin 합성경로의 중요 효소의 하나인 DFR 유전자를 분리하였다. 염기서열분석을 수행하여 분리유전자의 크기가 1279 bp이며 이 중 coding region은 1063 bp임을 확인하였다. 이미 밝혀진 다른 식물체의 DFR 유전자와 서로 염기서열의 일치성을 비교해 본 결과 Cheddar pink, 카네이션, 양배추, 개나리, 페튜니아, cup flower, 장미, 과꽃 및 거베라에서 각각 62% 이상을 나타내었다. 분리유전자의 발현을 확인하기 위하여 Northern blot 분석 및 인위적으로 기내에서의 transcription과 translation을 수행하였고, 분리한 유전자의 효소활성을 측정해 본 결과leucopelargonidin의 작은 peak를 확인하였다. Southern blot 분석 결과 안개초의 DFR 유전자는 다른 대부분의 식물체와 유사하게 한 개가 존재함을 확인하였다.

Agrobacterium-Mediated Transformation of Cucumber (Cucumis sativus L.) with Human BCL-2-encoding Gene

민병환,민병훈 한국원예학회 2009 Horticulture, Environment, and Biotechnology Vol.50 No.6

Transgenic cucumber (Cucumis sativus L.) plants were successfully obtained from cotyledon explants inoculated with Agrobacterium tumefaciens, which harbored a binary vector plasmid pPTN161 containing nptII gene for resistance to kanamycin and BCL-2 gene related to apoptosis from human. To establish high frequency of transformation system for cucumber, various conditions for transformation were investigated. After co-cultivation for 48 hours, the cotyledon explants were placed on shoot induction media (MS, 3 mg・L-1 zeatin, 0.3 mg・L-1 IAA) containing 100 mg・L-1 kanamycin and 300 mg・L-1 lillacilin. Shoot induction was continued for 3-4 weeks, then subcultured once, and after 2 weeks the shoots were transferred to root induction medium. After culture and selection on MS medium supplemented with 3 mg・L-1 zeatin, 0.3 mg・L-1 IAA and 100 mg・L-1 kanamycin, a number of kanamycin-resistant plantlets were regenerated. Polymerase chain reaction, Southern blotting analyses were used to identify and characterize the transgenic plants with the integrated human BCL-2 gene. Over 10 transgenic plants have been established in soil and flowered in the greenhouse. This procedure facilitates introduction of another desirable gene into commercial inbred lines of cucumber.

포도에 발생하는 해충의 종류와 무농약의 길

민병환 충남대학교 농과대학 2003 논문집 Vol.- No.9

Enterobacter agglomerans 339에 있어서 transposon umtagenesis를 통한 Nif$^{-10}$ -mutants 분리 동정

민병환,이호자 한국미생물학회 1988 미생물학회지 Vol.26 No.1

Three $NIf^{-}$ -mutants were isolated from Enterbacter agglomerans 339 through the transposon umtagenesis using a RP4-mobilising system for its nif-gene characterization. All mutants hadn't acetylene-reduction ability. Then we confirmed that Tn5 was inserted into all conserved nif-plasmids through the Southern Hybridization.