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Anti-sense RNA에 의한 C형 간염 바이러스의 번역의 억제
지승완,유병제 大邱大學校附設 基礎科學硏究所 1998 基礎科學硏究 Vol.14 No.2
The gene therpys have been investigated as the new therpy of the desease, the inhibition of the specific gene expressin by anti-sense RNA is an one method of the them. The inhibition of viral translation or viral replication by anti-sense RNA causes the viral proliferation. Since hepatitis C virus is RNA virus, HCV may be the target virus of anti-sense RNA as anti-viral reagent. The inhibition of the translation of the defective virus having CAT as the reporting gene by anti-sense RNA was performed. The inhibition of CAT translation did not occure in cells pretransfected with anti-sense RNA, the inhibition of CAT tranalation occured in cells co-transfected with anti-sense RNA. It seems that the difference between pretransfection and co-transfection was resulted from the cellular delivery system of anti-sense.
pT7T7을 이용한 인간 세포에서의 외래성 유전자 발현
지승완,유병제 大邱大學校附設 基礎科學硏究所 1998 基礎科學硏究 Vol.14 No.2
The viral cell culture is essential for the research of hepatitis C virus, but the cell culture of HCV has been not established recently. Since the cell culture by the RNA transfection has the problems in the quantifiction of transfected RNA, the cell culture by the DNA transfection has been tried. The expression of CAT as reporting gene by pT7T7 transfection in Huh7 cells was performed as the preexperiment of the HCV cell culture by the DNA transfection. As increasing of the concentration of target gene DNA, the expression of CAT was increased. The CAT was more expressed in the cells transfected with EMCV-CAT than in the cells with HCV-CAT. The difference between EMCV and HCV was resulted from the ability of enhancement of translation in untranslational region used for regulatory site of virus.
지승완,유병제 大邱大學校附設 基礎科學硏究所 1998 基礎科學硏究 Vol.14 No.2
Generally, the detection of virus was performed at the level of viral proteins and viral genome. The detection of virus by the viral genome more directly identifies the virus and estimates the concentration of virus. Because the genome of hepatitis C virus is RNA, the detection of the HCV genome was done by RT-PCR. But, PCR has the problems in contamination and false positive result. To deduce the upper problems, we developed the RT-PCR of the two ends of the HCV genome by long range cDNA. It seems that this method deduce the problems of contamination and false positive result, identify the full length genome of HCV and estimate the accurate concentration of HCV, and the fidelity and sensitivity of this method are good.
인간 간세포에서 C형 간염 바이러스의 replicase발현
지승완,유병제 대구대학교 기초과학연구소 1998 基礎科學硏究 Vol.15 No.1
To analyze the RdRp in eukaryotic HuH7 cells transfected defective HCV RNAs, cotransfection of defective HCV RNAs and defective HCV-CAT negative RNAs that had the essential portion of HCV for replication and translation, and CAT as reporting gene was performed. In cells transfected with defective HCV RNA and negative HCV-CAT RNA, the amount of CAT was quantified by Chloramphenicol acetyl transferase(CAT) enzyme-linked immunosorbent assay(ELISA). The NS5b of HCV was expressed from defective HCV 14H RNA and had a activity of RdRp. And, the defective HCV RNA and defective negative HCV-CAT RNA were replicated without other viral factors.
지승완,유병제 大邱大學校附設 基礎科學硏究所 1998 基礎科學硏究 Vol.15 No.1
The cells transfected with defective HCV RNAs alone, were harvested at various days after transfection. The replication of various defective HCV RNAs was checked by the quantification of HCV RNAs in transfected cell. The amount of HCV RNAs were estimated by the chiron bDNA kit. All the case, the amounts of HCV RNAs in transfected cells were decreased.
지승완,유병제 大邱大學校附設 基礎科學硏究所 1998 基礎科學硏究 Vol.15 No.1
Hepatitis C virus (HCV) is the major causative agent of posttransfusion and sporadic non-A, non-B hepatitis. To time, many research for HCV have been performed, but the molecular mechanism of HCV replication is still not obvious. To define the essential region of HCV genome for replication and develop the RNA-dependent RNA polymerase(RdRp) of HCV assay system in vivo. I constructed defective HCV cDNA that contained 5'Untranslate reagion,(UTR), Core(C), Enveloper 1(4 amino adds), Nonstructure(NS)5b, and various 3'UTR.
지승완(Seungwan Jee),김창환(Changhwan Kim),박미선(Misun Park),엄미옥(Miok Eom),염태경(Taikyung Ryeom),김옥희(Okhee Kim),강호일(Hoil Kang) 한국독성학회 2005 Toxicological Research Vol.21 No.1
Chrysin (5,7-dihydroxyflavone) is a flavonoid compound contained in many fruits, vegetables and honey. In our experiment, we investigated genotoxicity of chrysin using bacterial reverse mutation assay, chromosomal aberration test, in vivo micronucleus test. In bacterial reverse mutation assay, chrysin did not induce mutagenicity in Salmonella typhimurium TA98, TA100, TA1535, TA1537, TA102 with and without metabolic activation. In chromosome aberration test, chrysin did not also induce structural and numerical abberations regardless of metabolic activation in Chinese hamster lung fibroblast cells. In mouse micronucleus test, no significant increase in the occurrence of micronucleated polychromatic erythrocytes (MNPCE) was observed in ICR male mice orally administered with chrysin at the dose of 0.5, 1.0, 2.0 g/kg body weight. Taken together these results, chrysin has no mutagenic potential in our experiment.
알츠하이머 형질전환 마우스의 출생 후 뇌에서 유전자 발현
지승완(Seung Wan Jee),송연숙(Youn Suk Song),오재호(Jae Ho Oh),김용규(Yong Kyu Kim),심선보(Sun Bo Shim),황대연(Dae Youn Hwang),이수해(Su Hae Lee),서수진(Su Jin Seo),조준용(Jun Yong Cho),조정식(Jung Sik Cho) 한국실험동물학회 2005 Laboratory Animal Research Vol.21 No.3
DNA microarrays are a well-established technology for measuring gene expression levels. The aim in the study was to gain insight into potential over-expressed effect of amyloid precursor protein sw (APPsw) on modulation of genes for Alzheimer's Disease (AD). APPsw-transgenic mice, which has been produced by us previously, provide an important resource for identifying differentially expressed genes, since this transgenic line was shown cognitive deficits along with Aβ-42 depositions at 12 months of age. Using cDNA microarray, we compared mRNA levels in 7.4K cDNA clones from postnatal brain of age-matched wild-type mice and Alzheimer's diseased transgenic mice. A total of 44 differentially expressed genes with a 15 up-regulated and with a 29 down-regulated were found in brains from moderatively transgenic mice compared to non-transgenic littermates. Thus, the results allow to the future studies in the functions of each gene, which may be targeted for developing drugs.