• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.113-113
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• 1993

항바이러스 작용을 기대하여 6-azauridine의 2',3'-seco 유도체와 이들의 cyclic phosphate 유도체를 합성하였다. 6-azauridine으로부터 periodate oxidation에 의해 2',3'위치를 개열, 환원하여 얻은 triol의 3',5'위치를 acetonide 형으로 보호한 후 2'위치를 tosylation, azidation 하고 deprotection하여 2'-azido-2',3'-seco 유도체 (1)를 얻었다. 2',3'-diazido-2',3'-seco 유도체(2)는 6-azauridine의 2',3'-위치를 먼저 보호한후 5'위치를 benzoylation, 2',3'-deprotection, periodate oxidation 개열로 얻은 diol을 tosyl화 azido화 하고 deprotection하여 얻었다. 5'-azido-2',3'-seco 유도체(3)는 화합물 (2)의 합성시와 동일하게 먼저 2',3'-acetonide로 한후 5'-위치를 tosyl화, azido화 하고 deprotection, 2',3'-perodate 산화 개열등으로 얻었다. cyclic phosphate 유도체 (4)는 상기화합물 (2)를 4-NPPDC로 처리하여 3',5'-p-nitrophenylphosphoryltrioxy 유도체 (5)를 얻고 이것으로부터 3',5'-phosphoryltrioxy 유도체의 ammonium salt(4)를 얻었다. 이들 화합물의 DNA 및 RNA virus에 대한 antiviral activity는 현재 수행중이다.

• Korean Journal of Microbiology
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• v.24 no.3
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• pp.276-287
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• 1986

Chlorella ellipsoidea were cultured in the media containing rifampicin for 7 days. Aliquot cells were taken out after the inoculation and at intervals during cultivation and growth rate of Chlorella cells was measured. In order to investigate the effect of rifampicin on the nucleic acid synthesis, nucleic acid and RNA polymerase were extracted from chloroplast isolated from these cells, and the contents of nucleic acid and activity of enzyme were measured to compared with those of the control. The inhibitory concentration of rifampicin on growth was 80 ppm. The DNA contents in chloroplasts isolated were decreased 60% to compared with control, whole cells were markedly decreased 70% by rifampicin. The contents of base in the RNA were decreased 46% by rifampicin in shole cell, and 77% of base contents were decreased in chloroplast. Rifampicin also inhibited the activity of RNA polymerase, therefore whole cell was decreased 10% of activity and chloroplasts were decreased 42% of activity.

• Journal of Life Science
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• v.23 no.12
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• pp.1428-1435
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• 2013

The aim of this study is to investigate the melanogenic effect of Oenanthe javanica ethanolic extracts (OJE) containing quercetin and kaempferol in melanoma cells (B16F1). In order to determine whether OJE inhibits melanin synthesis at the cellular level, the melanoma cells were cultured in the presence of different concentrations of OJE. In the present study, the antioxidant effects of OJE on DPPH radical scavenging, power reduction, lipid peroxidation, and DNA oxidation were evaluated in a cell free system. Furthermore, the effect of OJE on the production of melanin was determined by dopaquinone (DOPA) assay and tyrosinase activity. In addition, the protein expression of tyrosinase, as well as antioxidant enzymes such as superoxide dismutase (SOD)-1, SOD-2 and glutathione reductase (GSH), were examined using Western blot analysis. In this study, it was observed that OJE exhibited an inhibitory effect on lipid peroxidation and blocked the DNA oxidation induced by the hydroxyl radical produced by Fenton's reagent. OJE increased melanin synthesis above 50 ${\mu}g/ml$ and tyrosinase activity was detected above 50 ${\mu}g/ml$. In Western blot analysis, OJE increased the expression levels of tyrosinase, SOD-1, SOD-2, and GSH in a dose-dependent manner. These findings indicate that OJE with antioxidant activity can regulate the tyrosinase activity and melanin production in melanocyte, suggesting that it could promote the development of black hair as well as protect skin from oxidative stress.

• The Korean Journal of Pesticide Science
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• v.18 no.3
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• pp.202-209
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• 2014

RNA interference (RNAi) is the method which controls phenotypes of gene in live cells. Chitinase is the enzyme helping digestion and absorption of old cuticles during the ecdysis of insects. In order to investigate molting-inhibition effect with the chitinase related gene in Spodoptera litura, RNA was extracted from the $5^{th}$ instars. cDNA was synthesized and then we obtained about 700 bp size chitinase. After PCR products were cloned into a pGEM T-easy vector, colonies were picked. DNA was extracted from the colony cultures. EcoR I enzyme was used to check whether PCR products were inserted or not. And then we confirmed vector band of about 3 kb and insert band of about 700 bp. To synthesize the dsRNA, each DNA was cut with Spe I and Nco I enzymes (Circular DNA became lineared DNA). After synthesis of dsRNA, approximately 5 ul dsRNA was injected into the $3^{rd}$ abdominal segment of S. litura $4^{th}$ larvae. The concentration of dsRNA was about $10{\mu}g/{\mu}l$. We confirmed larval-larval molting : there were phenotypically abnormal individuals - for instance malformation, molting inhibition and change of integument color. Pupaadult molting : there were phenotypically abnormal individuals - for instance molting inhibition, change of wings and malformation. Also we could investigate the pupation, emergence and variation about noninjection, treated with DW and dsRNA. Each pupation was non-injection 83.3%, DW 78.3% and dsRNA 66.7%. Each emergence was non-injection 90.0%, DW 72.3% and dsRNA 65.0%. So we considered that chitinase dsRNA induced molting inhibition effect. But each variation was non-injection 8.9%, DW 2.9% and dsRNA 19.2%. Therefore dsRNA group showed the highest variation value. When 18 hours after injecting dsRNA, we could obtain abnormal individual.

• Journal of Food Hygiene and Safety
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• v.28 no.2
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• pp.181-187
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• 2013

Identification of main ingredients in starches has been investigated using physicochemical analysis method mainly. However, physicochemical properties such as particle size have limitations in determining the differences among mixed starches. Therefore, we developed a molecular biological method to identify materials used in starch, as a sample, 11 kinds of starches including sweet potato starch, potato starch, corn starch, and tapioca starch. DNeasy plant mini kit, magnetic DNA purification system, and CTAB methods were used to extract DNA from samples. After gene extraction, whole genome amplification (WGA) was performed to amplify the extracted DNA. Species-specific primers were used as followings: ib-286-F/ib-286-R (105 bp), Pss 01n-5'/Pss 01n-3' (216 bp), SS11b 3-5'/SS11b 3-3' (114 bp), and SSRY26-F/SSRY26-R (121 bp) gene for sweet potato, potato, corn, and tapioca, respectively. In this study, we could confirm the main ingredients using WGA and PCR method.

• Korean Journal of Plant Tissue Culture
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• v.25 no.5
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• pp.341-346
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• 1998

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.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.857-862
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• 2005

This study investigated the proliferation and DNA synthesis inhibitory effect of concentrations ($0.01\%$, $0.1\%$, $0.25\%$, $0.5\%$) when added whole molecular-, 40 kDa-, or 10 kDa polymannuronate on human colon cancer cells, HT-29, DLD-1, and WiDr, in vitro. In order to determine the proliferation inhibitory effect of low-molecularized polymannuronate, the treatment of whole molecular-, 40 kDa-, 10 kDa-, polymannuronate ($0.25\%$) to the HT-29 cancer cells inhibited proliferation of cancer cells by $41\%$, $69.1\%$, and $75.6\%$, respectively. DLD-1 cancer cell was not relation of molecular weight and concentration. WiDr cancer cell depend on concentration without molecular weight. In addition, whole molecular-, 40 kDa-, 10 kDa poly mannuronate ($0.25\%$) significantly inhibited DNA synthesis of HT-29 cancer .cells by $78\%$, $58\%$, and $56\%$, respectively. And morphological changes not found under microscope by polymannuronate. Therefore polymannuronate would be helpful to colon cancer treatment as well as cancer prevention and this study would be the basic source for further research of polymannuronate.

• Journal of Plant Biology
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• v.36 no.4
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• pp.415-423
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• 1993

대두의 uricase II cDNA를 탐침으로 plaque 혼성화 방법에 의해 해녀콩의 뿌리를 cDNA library로부터의 두 개의 phage 클론(λCINUO-01, λCINUO-02)을 선별하였다. 두 phage 클론은 약 1.6 kb와 1.0 kb의 insert를 갖고 있었으며 이들의 염기서열을 결정하기 위하여 pUC19과 pBSKS vector에 subcloing(pcCLNUO-01, pcCLNUO-02)하였다. Sanger법에 의해 염기서열을 결정한 결과, 두 클론은 각각 1,611 bp와 1,024 bp로 이루어져 있었으며 pcCINUO-01은 308개의 아미노산, pcCINUO-02는 301개의 아미노산을 암호화하는 open reading frame(ORF)을 갖고 있었다. 두 클론의 ORF의 염기서열은 대두의 uricase II와 각각 88.9%, 89.3%의 상동성을 보여주었으며, 아미노산 서열은 84.1%, 85.4%의 상동성을 보여주었다. pcCINUO-01의 경우, 종결코돈으로부터 313 NT 하류쪽에 진핵생물의 poly(A) 첨가신호인 AATAAA 서열이 존재하였으며 이로부터 21 NT 하류쪽에 17 잔기의 poly(A)가 존재하였다. 두 클론의 염기서열에서 추정된 아미노산 서열의 카르복시 말단에는 세포질에서 합성된 몇몇 단백질들이 peroxisome으로 수송되는데 필요한 신호서열인 Ser-Lys-Leu-COOH 서열이 존재하고 있었다. 두 클론의 염기서열을 토대로 아미노산 조성을 살펴본 결과, 염기성 아미노산(Arg, His, Lys)과 산성 아미노산(Asp, Glu)이 각각 46 대 35, 47 대 35의 비를 보여주었는데 이는 uricase II 단백질의 염기성 성질을 보여주는 결과로 추정된다. Northern 혼성화 결과 해녀콩에서 uricase II는 뿌리혹에서만 특이적으로 발현됨을 알 수 있었고 게놈 혼성화 반응 결과는 uricase II 유전자가 해녀콩 게놈상에 유전자 가족으로는 존재할 수 있음을 보여주었다.

• Applied Biological Chemistry
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• v.40 no.1
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• pp.30-33
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• 1997

About 250 bp ura5 gene (Orotate phosphoribosyl transferase) fragment was cloned from genomic DNA of entomopathogenic fungus Metarhizium anisopliae by using PCR method. Entire nucleotide sequences of cloned DNA fragment were determined and analysed as compared with other fungus ura5 genes. The amino acid sequence deduced from the nucleotide sequence showed 85.5% homology to ura5 protein of Trichoderma reesei. Using this 250 bp PCR fragment we have isolated full ura5 gene of M. anisopliae by genomic Southern hybridization and the isolated 4.4 kb DNA fragments were mapped by restrictional enzyme.

• YAKHAK HOEJI
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• v.39 no.6
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• pp.661-665
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• 1995

Ginsenoside-Rg$_{1}$(G-Rg$_{1}$) has been shown to stimulate DNA synthetic activity in SK-HEP-1 cells. This study was therefore designed to determine in SK-HEP-1 cells whether the stimulatory effect of G-Rg$_{1}$ may be mediated by protein kinase C (PKC) which is known to play a key role in the signal transduction pathway leading to the cell proliferation. Using the tn situ PKC assay method, the PKC enzyme activity was determined in SK-HEP-1 cell cultures in response to G-Rg$_{1}$ at 3*10$^{-5}$ M or phorbol 12-myristate 13-acetate(PMA) at 10$^{-6}$ M which in the enzyme activity by 1.5- and 7-fold, respectively. Furthermore, G-Rg$_{1}$, was also able to synergistically increase the enzyme activity by 11-fold m the cell cultures in the presence of PMA. These stimulatory effects of G-Rg$_{1}$ or PMA on the DNA synthetic activity and the PKC activity were ablished by a specific PKC inhibitor, GF109203X. These results suggest that the stimulatory effect of G-Rg$_{1}$ on the DNA synthetic activity may be partly due to stimulation of PKC-mediated signal transduction pathway leading to the proliferation of SK-HEP-1 cells.