• 대한수혈학회지
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• 제23권2호
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• pp.152-161
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• 2012

배경: 2012년 1월부터 국내에 신규로 도입되는 고위험군 바이러스 검사시약은 국내 검체를 이용한 성능 시험분석을 실시하여 식품의약품안전청 허가를 득하도록 일부 규정이 개정되었다. 이러한 바이러스 진단시약의 성능시험과 품질관리 규정을 이행하기 위해서는 국내 검체에서 유래된 국가표준물질이 필요하다. 이에 본 연구에서는 바이러스 핵산증폭검사시약의 성능평가와 품질관리를 위하여 다양한 농도로 구성된 HBV, HCV와 HIV의 핵산증폭검사용 혈장유래 표준물질(plasma working standards)을 제조하고자 하였다. 방법: 수혈 부적격의 HCV RNA 양성혈장 43단위, HCV RNA 양성혈장 25단위, 그리고 HIV RNA 양성혈장 26단위에 대해 핵산 정량검사와 유전자형 검사를 실시하였다. 이를 바탕으로 국내에서 유행하는 바이러스의 유전자형을 가진 고농도의 원료혈장을 선정하였다. 또 표준물질의 다양한 농도는 국내외 검사시약의 검출범위에 근거하여 적정한 농도범위를 선택한 후, 다양한 농도로 원료물질을 희석하고, 분병 처리 한 후 정량적으로 분석하였다. 결과: HBV, HCV와 HIV의 체외진단분석기용 핵산증폭검사의 성능평가용 plasma working standards는 총 13종의 다양한 농도물질로 제조되었다. 결론: 국내에서 혈액관련 고위험군 HBV, HCV와 HIV의 체외진단용 핵산증폭검사 시약의 성능 평가에 필요한 바이러스 핵산 검사용 국가표준물질을 처음 제조하여 그 기술을 수립하였다.

• 공업화학
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• 제10권4호
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• pp.599-602
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• 1999

Chitosan을 아질산부해법을 이용하여 저분자화시킨 다음, 저분자량 chitosan의 유리 아민기와 trifluoroacetic anhydride를 반응시켜 새로운 함불소 chitosan 올리고머 유도체(FCO)를 합성하였다. 이들 반응의 진행은 FT-IR, $^{1}H\;NMR$, $^{19}F\{^{1}H\}NMR$ 등을 통해 확인하였다. FCO의 항바이러스 효과는 바이러스 감염용액에 다양한 농도의 FCO를 첨가하고 세포를 감염시킨 후, 36시간 후에 복제되고 있는 바이러스 DNA 양을 측정하여 조사하였다. 바이러스 복제는 FCO를 첨가하여 바이러스를 감염시킨 세포들에서, FCO를 첨가하지 않은 대조군에 비하여 첨가한 FCO의 농도에 비례하여 감소하여, FCO가 바이러스 감염을 효과적으로 억제함을 제시한다. 특히, 1% FCO의 감염용액으로 처리된 세포에서는 바이러스의 복제가 대조군에 비하여 40%로 감소하였다.

• 대한수의학회지
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• 제36권4호
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• pp.859-871
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• 1996

This study was conducted to elucidate the distribution of Aujeszky's disease viral nucleic acids and antigens in the central nervous system (CNS) of piglets. The first Korean isolate of Aujeszky's disease virus(ADV) that isolated from naturally infected piglets in Yang San, was inoculated into 32 day old piglets with $10^{5.9}TCID_{50}/ml$ through intranasal or intramuscular route. These piglets were sacrificed at every 24hrs for 8 days. The immunohistochemistry (IHC) was conducted to detect the viral antigens in paraffin-embedded tissue sections using avidin-biotin-peroxidase complex (ABC) method. The viral nucleic acids were detected by in situ hybridization (ISH) using ADV specific DNA probe labeled with digoxigenin. The ADV antigens were detected in reticuloendothelial cells of spleen, lymph nodes and tonsil, alveolar walls, leptomeningeal vascular walls, inflammatory foci of each organ, and nerve cells. The viral nucleic acids were detected in the spinal trigeminal nucleus and its tracts of the pons and medulla oblongata by the ISH technique. The pathways of AD viruses in CNS were determined by IHC and ISH. In the intranasally inoculated group, the viruses in nasal mucosa moved to medulla oblongata and pons through the trigeminal nerve. In case of intramuscullarly inoculated group, viruses moved to brain via lymphoid organs or spinal nerves from sciatic nerves.

• 한국어병학회지
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• 제35권2호
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• pp.141-155
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• 2022

Rock bream iridovirus (RBIV) causes high mortality and economic losses in the rock bream (Oplegnathus fasciatus) aquaculture industry in Korea. Viral open reading frames (ORFs) expression profiling at different RBIV infection stages was investigated using microarray approaches. Rock bream were exposed to the virus and held for 7 days at 23 ℃ before the water temperature was reduced to 17 ℃. Herein, 28% mortality was observed from 24 to 35 days post infection (dpi), after which no mortality was observed until 70 dpi (end of the experiment). A total of 27 ORFs were significantly up- or down-regulated after RBIV infection. In RBIV-infected rock bream, four viral genes were expressed after 2 dpi. Most RBIV ORFs (26 genes, 96.2%) were significantly elevated between 7 and 20 dpi. Among them, 12 ORF (44.4%) transcripts reached their peak expression intensity at 15 dpi, and 14 ORFs (51.8%) were at peak expression intensity at 20 dpi. Expression levels began to decrease after 25 dpi, and 92.6% of ORFs (25 genes) were expressed below 1-fold at 70 dpi. From the microarray data, in addition to the viral infection, viral gene expression profiles were categorized into three infection stages, namely, early (2 dpi), middle (7 to 20 dpi), and recovery (25 and 70 dpi). RBIV ORFs 009R, 023R, 032L, 049L, and 056L were remarkably expressed during RBIV infection. Furthermore, six ORFs (001L, 013R, 052L, 053L, 058L, and 061L) were significantly expressed only at 20 dpi. To verify the cDNA microarray data, we performed quantitative real-time PCR, and the results were similar to that of the microarray. Our results provide novel observations on broader RBIV gene expression at different stages of infection and the development of control strategies against RBIV infection.

• BMB Reports
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• 제31권3호
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• pp.303-306
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• 1998

Serine proteinase cDNA fragment from protozoan parasite Acanthamoeba culbertsoni was amplified by the reverse transcription-polymerase chain reaction (RTPCR) using degenerate oligonucleotide primers derived from conserved serine proteinase sequences. The amplified DNA fragment was subcloned and sequenced. The sequence analysis and alignment showed significant sequence similarity to other eukaryotic serine proteinases and conservation of the His, Asp, and Ser residues that form the catalytic triad. The cDNA fragment was cloned into the pGEMEX-1 expression vector and expressed in Escherichia coli. A resulting fusion protein of 56 kDa had proteolytic activity. The fusion protein reacted with sera of mice immunized with purified serine proteinase of A. culbertsoni in Western blot. Immune recognition of the fusion protein by mouse antisera suggested that the fusion protein may be valuable as a diagnostic reagent.

• 한국수의병리학회지
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• 제7권1호
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• pp.1-4
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• 2003

Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by a gamma herpesvirus, ovine herpesvirus 2 (OvHV-2). Dot blot hybridization (DBH) protocols for detecting and differentiating this MCF virus were developed. OvHV-2 specific primer pairs, 556/555, were used for the amplification of target DNA. Then, the amplified DNA was labeled with incorporation of digoxigenin (DIG). The Dig-labeled probe was able to detect and differentiate specifically OvHV-2 DNA. This DBH technique can be applied to confirm the presence of MCF virus on clinical samples and to differentiate specifically between OvHV-2 infection and other viral infections.

• 한국식물병리학회지
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• 제11권1호
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• pp.87-90
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• 1995

Complementary DNA of the movement protein (MP) gene of tobacco mosaic virus Korean pepper strain (TMV-KP) was synthesized from purified TMV-KP RNA by using the reverse transcription and polymerase chain reaction (PCR) system. The synthesized double stranded cDNA was cloned into the plasmid pUC9 and transformed into Escherichia coli JM110. The movement protein gene of TMV-KP of the selected clones was subjected to sequence analysis by Sanger's dideoxy chain termination method. The complete sequence of viral MP gene from TMV-KP strain was 807 nucleotides long. The nucleotide of MP gene from TMV-KP has thirteen and two nucleotide differences from TMV vulgarae (TMV-OM) and Korean (TMV-K) strains, respectively. Thus, the nucleotide sequence of TMV-KP MP gene showed higher homology of 99% with that of TMV-K MP gene.

• 한국연초학회지
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• 제23권2호
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• pp.103-108
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• 2001

Hog cholera virus(HCV) was purified from virus infected Bovine kidney cells. From this virus, total protein was analyzed by SDS-PAGE gel electrophoresis and about 55 kDa band of E2 envelope protein was detected. The viral RNA was purified and E2 cDNA was amplified by RT-PCR. E2 cDNA fragment was cloned to PCRII-TOPO cloning vector and named pE2. The analysis of nucleotide sequence showed that this E2 cDNA fragment inserted into pE2 was 1191 nucleotides long and coded 397 amino acids.

• Journal of Plant Biotechnology
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• 제47권2호
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• pp.164-171
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• 2020

Transgenic Alstroemeria plants resistant to Alstroemeria mosaic virus (AlMV) were generated through RNA-mediated resistance. To this end, the friable embryogenic callus (FEC) of Alstroemeria was induced from the leaf axil tissue and transformed with a DNA fragment containing the coat protein gene and 3'-nontranslated region of AlMV through an improved particle bombardment system. The bar gene was used as a selection marker. More than 300 independent transgenic FEC lines were obtained. Among these, 155 lines resistant to phosphinothricin (PPT) were selected under low stringent conditions. After increasing the stringency of PPT selection, 44 transgenic lines remained, and 710 somatic embryos from these lines germinated and developed into shoots. These transgenic shoots were then transferred to the greenhouse and challenged with AlMV. In total, 25 of the 44 lines showed some degree of resistance. PCR analysis confirmed the presence of the viral sequence. Virus resistance was observed at various levels. Establishment of an efficient transformation system for Alstroemeria will allow inserting transgenes into this plant to confer resistance to viral and fungal pathogens. Accordingly, this is the first report on the production of a transgenic virus-resistant Alstroemeria and lays the foundation for alternative management of viral diseases in this plant.

• 대한한방종양학회지
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• 제4권1호
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• pp.199-209
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• 1998

In order to find antiviral effect against Human immunodeficiency virus(HIV), Herpes simplex virus type I(HSV-1) and II(HSV-2) from herb medicines, publicated 29 paters on anti-viral effect of herbal medicines and a convenient virus-induced cytopathic effect (CEP) inhibition assay was introduced. The major virus on experiment are HIV, Hepatitis B virus and HSV-1,2. Those of other studies showed inhibition of infected virus DNA replication and screening test of herbal medicines. More than 15 extractions were prepared by pure water boiling from herbal medicines, and their toxicity of infected cell and anti-viral activities were evaluated. Among them, the major part of herbal medicines showed cell stability compared with the contrast. Cytotoxic concentration (CC) of the $H_2O$ extracts of Padoo against HIV was <4.0, Hyungbangpaedoksan against HIV was 9.3, Whangyonhaedoktang against HIV-1 and HSV-2 was 15.3. These are high level cytotoxic concentration compared with the contrast. But antiviral effect was unable to figure out for selective $index(SI)=CC_{50}/EC_{50}$. The other herbal medicines were unable to showed potent anti-HIV and anti-HSV activity. The antiviral activation using herbs in this thesis have unlimited objects, to select research object will help to show the direction of antiviral drug development that have less side effect and more excellent efficiency.