지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2019.6
- 수록면
- 149 - 156 (8page)
- DOI
- 10.17519/apiculture.2019.06.34.2.149
이용수
초록· 키워드
오류제보하기
In Korea, sacbrood virus infecting Apis cerana (AcSBV) is a small RNA virus belonging to the family Iflaviridae. AcSBV causes larval death, and even the collapse of entire bee colonies and its viral capsid is composed of four structural proteins (VP1, VP2, VP3 and VP4). In this study, we synthesized VP3 gene based on genomic sequences to insert into a His-tag expression plasmid of pET-28a. The resultant recombinant vector was then transformed into Escherichia coli BL21 (DE3) to express the recombinant VP3 protein (rVP3). The rVP3 protein was strongly expressed in an insoluble fraction, with a molecular size of around 26 kDa. Purification of the rVP3 protein using an affinity Ni-NTA resin, followed by SDS-PAGE separation, resulted in a high purity of the protein and used as an antigen to raise the polyclonal antibodies. Mice were then immunized with the purified rVP3 protein and the antiserum collected at 4<SUP>th</SUP> week and 6<SUP>th</SUP> week was able to detect the rVP3 at a concentration of 500 ng and 250 ng. Furthermore, IgG antibody levels reacted with wild-type larva crude protein extract, AcSBV-infected larva crude protein extracts and rVP3 protein were quantitatively analyzed by ELISA and qualitatively analyzed by western blotting. This is the first report on production of polyclonal antibodies directed against the rVP3 protein of AcSBV and its use for detection and diagnosis of virus from field samples.
목차
Abstract
INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
LITERATURE CITED
참고문헌 (36)
참고문헌 신청
Ai, H. / 2012 / Occurrence and prevalence of seven bee viruses in Apis mellifera and Apis cerana apiaries in China / J. Invertebr. Pathol 109:160~164
Aruna, R. / 2018 / Complete genome sequence of sacbrrod virus isolated from Asiatic honey bee Apis cerana indica in India / Virus Dis 28:453~460
Butt, T. R. / 2005 / SUMO fusion technology for difficult-to-express proteins / Protein Expre. Purif 43:1~9
Chen, Y. / 2004 / Multiple virus infections in the honey bee and genome divergence of honey bee viruses / J. Invertebr. Pathol 87:84~93
Choe, S. E. / 2012 / Analysis of the complete genome sequence of two Korean Sacbrood viruses in the honey bee, Apis mellifera / Virology 432:155~161
Aruna, R. / 2018 / Complete genome sequence of sacbrrod virus isolated from Asiatic honey bee Apis cerana indica in India / Virus Dis 28:453~460
Butt, T. R. / 2005 / SUMO fusion technology for difficult-to-express proteins / Protein Expre. Purif 43:1~9
Chen, Y. / 2004 / Multiple virus infections in the honey bee and genome divergence of honey bee viruses / J. Invertebr. Pathol 87:84~93
Choe, S. E. / 2012 / Analysis of the complete genome sequence of two Korean Sacbrood viruses in the honey bee, Apis mellifera / Virology 432:155~161
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UCI(KEPA) : I410-ECN-0101-2019-527-000882038