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        돼지 group C 로타바이러스 VP6 특이 단클론항체

        윤영심 ( Young Sim Yoon ),이승철 ( Seung Chul Lee ),우상규 ( Sang Kyu Woo ),조경오 ( Kyoung Oh Cho ),강신영 ( Shien Young Kang ) 한국동물위생학회 2012 韓國家畜衛生學會誌 Vol.35 No.3

        Rotaviruses have been known to be a major etiological agent of gastroenteritis in both infants and young animals. Subsequently new rotaviruses, which were morphologically indistinguishable but antigenically and electrophoretically distinct with each other, were reported from several animals throughout world including Korea. These new rotaviruses were named as non-group A or group B or group C rotaviruses and so on. It has been very difficult to isolate and grow the non-group A rotaviruses in vitro, and this has greatly limited the characterizations of non-group A rotaviruses and serological studies. In this study, monoclonal antibodies (MAbs) against porcine non-group A rotavirus were produced and characterized. The VP6 gene of porcine group C rotavirus Korean isolate(#06-52-1) was cloned and expressed. For expression of VP6 gene, baculovirus expression system was applied. The VP6 gene and expressed protein in the recombinant virus were confirmed by polymerase chain reaction (PCR), indirect fluorescence antibody (IFA) test and Western blot, respectively. The expressed VP6 was used for MAbs production. The MAbs produced in this study would be promising as diagnostic reagents for detection of group C rotavirus infection.

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        광견병바이러스에 대한 단크론항체 생산 및 특성

        이승철 ( Seung Chul Lee ),윤영심 ( Young Sim Yoon ),송윤경 ( Yun Kyung Song ),우계형 ( Gye Hyeong Woo ),진영화 ( Young Hwa Jean ),강신영 ( Shien Young Kang ) 한국가축위생학회 2010 韓國家畜衛生學會誌 Vol.33 No.2

        Rabies virus which belongs to the genus Lyssavirus of the family Rhabdoviridae is known as a highly neurotropic virus and causes fatal encephalitis accompanied by severe neurological symptoms in almost all mammals, including humans. In this study, monoclonal antibodies (MAbs) against rabies virus were produced, characterized and applications of MAbs as diagnostic reagents were assessed Spleen and inguinal lymph node cells from Balb/c mouse immunized with purified rabies virus were fused with SP2/O myeloma cells using polyethylene glycol (PEG) and hybridoma cells producing rabies virus-specific MAbs were screened by an indirect fluorescent antibody test. A total of ten MAbs were produced against rabies virus. The protein specificity and neutralizing activity of MAbs were determined by Western blot analysis and fluorescent antibody virus neutralization test, respectively. As a result, two MAbs, 5G3 and 6H4 had specificity for nucleoprotein (N protein) and two other MAbs, 5B1 and 5C1 had neutralizing activity for rabies virus. Some MAbs recognized the rabies virus-infected bovine brain stem cells by immunohistochemistry (IHC) assay. In conclusion, it was confirmed that MAbs produced in this study were rabies virusspecific and could be used as reliable diagnostic reagents for the detection of rabies virus.

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        돼지생식기호흡기증후군 바이러스의 ORF6 유전자 발현

        배수정 ( Su Jung Bae ),김진원 ( Jin Won Kim ),윤영심 ( Young Sim Yoon ),강신영 ( Shien Young Kang ) 한국가축위생학회 2009 韓國家畜衛生學會誌 Vol.32 No.1

        Porcine reproductive and respiratory syndrome (PRRS) virus is the etiological agent of diseases characterized by reproductive losses in sows and respiratory disorders in piglets. The PRRS virus is a small enveloped virus containing a positive-sense, single-stranded RNA genome. In the present study, ORF6 gene of Korean PRRS virus isolate, CNV, was cloned and expressed in baculovirus expression system. The ORF6 gene and expressed protein in the recombinant virus were confirmed by PCR/indirect fluorescence antibody (IFA) test and Western blotting, respectively. The recombinant protein with a molecular weight of approximately 24KDa was confirmed by Western blotting using His6 and PRRS virus-specific antiserum. Expressed ORF6 protein was applied for IFA to detect antibody against PRRS virus using field porcine sera. However, the sensitivity and specificity of developed IFA using expressed ORF6 protein were considerably low compared to those of commercial ELISA kit. This results suggest that IFA using expressed ORF6 protein could not be used as a diagnostic test for PRRS virus infection without further improvements.

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