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국내에서 분리된 소 로타바이러스의 VP7 유전자 크로닝 및 염기서열 분석
강신영,전성진,장경옥,박용하,김원용,Kang, Shien-young,Jeon, Seong-jin,Chang, Kyeong-ok,Park, Yong-ha,Kim, Won-yong 대한수의학회 1997 大韓獸醫學會誌 Vol.37 No.2
Bovine rotaviruses(A, 288, 55086 strains) isolated from fecal samples in Korea were propagated onto MA104 cells and were confirmed tentatively as G6, G8, and G10, respectively, by RFLP analysis. Full-length VP7 gene of these isolates was amplified by reverse transcriptase polymerase chain reaction(RT-PCR) using VP7 specific primers and cloned into TA vector. Nucleotide and deduced amino acid sequences of VP7 genes of the isolates were determined and compared with those of bovine rotavirus reference strains(NCDV; G6, UK; G6, Cody I-801; G8 and B223; G10). A, 288 and 55086 isolates showed high degree of nucleotide sequence homology with NCDV and UK(93% and 94%), Cody I-801(86%) and B223(97%), respectively, However, they showed 71~74% of nucleotide sequence homlogy with bovine rotavirus reference strains which belong to different serotypes. From the results of deduced amino acid sequence homology analysis, three isolates showed 94~96% of homology with the same serotype reference strains but 80~84% of homology with the different serotype reference strains. Three bovine rotavirus isolates, A, 288 and 55086 strains, were confirmed as G6, G8, and G10, respectively, by nucleotide and deduced amino acid sequence analysis.
Development of diagnostic methods for rotavirus from pigs using monoclonal antibody
강신영,Kang, Shien-young The Korean Society of Veterinary Science 1992 大韓獸醫學會誌 Vol.32 No.4
Group A 로타바이러스 VP6에 특이적으로 반응하는 단크론 항체를 이용하여 로타바이러스 감염이 의심되는 돼지 분변으로부터 로타 바이러스를 검색할 수 있는 효소면역측정법을 개발하였다. 이 효소면역측정법에서는 capture antibody로서 protein A-sepharose를 이용하여 단크론 항체로부터 순수 분리한 immunoglobulin을 사용하였으며 detecting antibody는 토끼 면역혈청으로부터 순수 분리한 immunoglobulin에 biotin을 label하여 사용하였다. 개발된 효소면역측정법의 민감도와 특이성을 전자현미경법 및 형광항체법의 것과 비교하여 보았을 때 서로 유사하였으며 분변재료로부터 로타바이러스를 검색하는데 유용한 것으로 나타났다. 개발된 효소면역측정법은 야외로부터 로타바이러스 검색을 위하여 수집된 많은 양의 분변재료를 실험실내에서 screen하는데 유용하게 사용될 것으로 생각된다.
안재문,강신영,Ahn, Jae-moon,Kang, Shien-young 대한수의학회 1998 大韓獸醫學會誌 Vol.38 No.3
Eight monoclonal antibodies(MAbs) against bovine coronavirus(BCV) were produced and characterized. Three MAbs(1G9, 4H12, 5C1) specific to the S glycoprotein and two HE glycoprotein-specific MAbs(2A5, 5G4) were found to neutralize the BCV in fluorescence focus neutralization(FFN) test. Two HE-specific MAbs from the neutralizing MAbs inhibited the hemagglutinating activity of the BCV. None of the N protein-specific MAbs(1C1, 5A12, 6H1) neutralized the virus infectivity. Bovine coronavirus and mouse hepatitis virus, which belong to group II coronaviruses, were differentiated from other groups of coronaviruses(porcine transmissible gastroenteritis virus, porcine epidemic diarrhea virus, canine coronavirus) by all MAbs in fluorescence antibody test(FA), but not in FFN test.
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 한국동물위생학회지 (KOJVS) 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.
소 로타바이러스(국내분리주)에 대한 단크론항체 생산 및 특성에 관한 연구
안재문,조선희,강신영,Ahn, Jae-moon,Cho, Sun-hee,Kang, Shien-young 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.2
Monoclonal antibodies(MAbs) against field isolates of the bovine rotavirus A strain(G6), V strain(G10) and reference I-801 strain(G8) were produced and characterized. Six MAbs(4C2, 4D9, 5E1, 5E7, 5D5, 3E4) against A strain had neutralizing activity and reacted only with the G6 bovine rotaviruses determined by fluorescence focus neutralization (FFN) test. Otherwise, five neutralizing MAbs(1G2, 2G6, 5E2, 5E12, 5H7) against I-801 strain neutralized the G6 and G8 bovine rotaviruses. Five non-neutralizing MAbs(5F12, 7F12, 5E11, 2A11, 2B12) were VP6-specific and cross-reacted with all bovine and porcine rotaviruses examined by fluorescence antibody(FA) test. None of the MAbs reacted with bovie viral diarrhea virus(BVDV) and bovine coronavirus(BCV) determined by FA and FFN test.
돼지 로타바이러스(Gottfried 주)의 VP4 항원구조분석
송윤경,김원용,강신영,Song, Yun-kyung,Kim, Won-yong,Kang, Shien-young 대한수의학회 2001 大韓獸醫學會誌 Vol.41 No.3
The neutralization epitopes of the outer capsid protein VP4 of a porcine rotavirus, Gottfried strain, were studied using neutralizing monocolonal antibodies(N-MAbs). Eight N-MAbs which are specific for the VP4 of Gottfried strain were used for analyzing the antigenic sites of VP4. Three different approaches were used for this analysis; i)testing the serological reactivity of each N-MAb against different G and P types of human and animal rotavirusese ii) analyzing N-MAb-resistant viral escape mutants and iii) performing nucleotide sequence analysis of the VP4 gene of each N-MAb-resistant viral escape mutant. From experimental results, at least four antigenic sites(I, II, III, and IV) were identified. Antigenic site I recognized by N-MAbs 24B9, 23G10, and 26A2 was separated from antigenic site II recognized by N-MAbs 30H5, 32B3, and 29B3. However, these antigenic sites were overlapped with antigenic site III recognized by N-MAb 21A1. The other antigenic site IV recognized by N-MAb 16D2 was separated from antigenic sites I, II, and III.
Serologic and electropherotypic characterization of the bovine rotaviruses isolated in Korea
정정원,장정호,강신영,박봉균,조재진,안수환,Chung, Chung-won,Chang, Chung-ho,Kang, Shien-young,Park, Bong-kyun,Cho, Jae-chin,An, Soo-hwan The Korean Society of Veterinary Science 1998 大韓獸醫學會誌 Vol.38 No.1
국내분리 소 로타바이러스의 genomic RNA 형태, 분리주간의 교차면역반응 그리고 단클론 항체를 이용한 중화시험에 의해 혈청형을 조사한 바 다음과 같은 결과를 얻었다. 국내분리주들의 genomic RNA 형태는 크게 NCDV 형태(7/11 시료)와 non-NCDV 형태(4/11 시료)의 두가지로 나타났다. 교차면역시험에서 표준주인 NCDV주에 대한 양성혈청은 국내분리주들에 대해서는 비교적 낮은 중화력을 나타내었으나 국내분리주에 대한 양성혈청들은 타 국내분리주 뿐만 아니라 NCDV주에 대해 서로 높은 중화력을 나타내었는데, 분리주 중 678, P44, M4에 대한 양성혈청은 NCDV를 포함한 대부분의 분리주에 대해 100%의 중화력을 나타내었다. 또한 288주에 대한 양성혈청은 288, 678,P44, M4주에 대해서는 높은 중화력을 나타내었으나 다른 분리주들에 대해서는 비교적 낮은 중화력을 나타내었다. 국내분리주의 단크론 항체를 이용한 G혈청형 감별결과는 G6유사형이 45.8%(11/24주), G10 유사형이 54.2%(13/24주)로서 두가지 G형이 존재하였다. 한편 G6형에 반응한 것들의 P형은 표준주인 NCDV주(P1)와는 다른 것으로 확인되었으며, G10 유사형에 속하는 14주는 모두 P11 혈청형으로 판명되었다.