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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2014.4
- 수록면
- 57 - 62 (6page)
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이 논문의 연구 히스토리 (2)
2014
Over-expression and Purification of Peptidase-C3G protein of Deformed Wing Virus in E. coli system
Joo-seong Lee
,
Hee-Young Lim
,
Byoung-Su Yoon
한국양봉학회 학술대회 자료집
2014.04
학술대회자료
Over-expression and Purification of Peptidase-C3G Protein of Deformed Wing Virus
Joo-seong Lee
,
Hee-Young Lim
,
Byoung-Su Yoon
Journal of Apiculture
2014.04
학술저널
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초록· 키워드
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Deformed wing virus (DWV) is associated with wing deformity of honeybee and is transmitted through Varroa destructor. In this study, recombinant peptidase-C3G that is one of the polyprotein of DWV was successfully over-expressed in E.coli system and purified by histag purification kit. Amplified peptidase-C3G fragment was constructed into expression vector pET32 by BamHI and SalI site and recombinant clone was named as pET32a-DWV-C3G. The peptidase-C3G in clone showed the 96% and 99% homology of nucleotide sequence and amino acid sequence of DWV complete genome (accession NO. JX878305.1), respectively. Recombinant DWV-C3G protein was optimized the expression under 0.6mM IPTG concentration for 6hr induction time at 25°C. The bacterial cells were harvested, lysed in PBS using sonication and then confirmed the overepxression of target protein in SDS PAGE. As a result, over-expressed DWV-C3G protein was observed around 27KDa and purified using His-tagging system based on high affinity chromatography between immobilized Ni-ion and histidine amino acid. This purified recombinant DWV-C3G protein will be used for the generation of monoclonal antibody to develop a diagnosis tool for DWV infection in honeybee.
목차
Abstract
INTRODUCTION
MATERIALS AND METHODS
RESULTS AND DISCUSSION
LITERATURE CITED
참고문헌 (12)
참고문헌 신청
1. [학술지(정기간행물)] - Bakonyi, T. / 2002 / Phylogenetic Analysis of Acute Bee Paralysis Virus Strains / Applied and Environmental Microbiology Dec 2002 : 6446 ~ 6450
2. [단행본] - Baily, L. / 1991 / Honey bee pathology / Academic Press
3. [학술지(정기간행물)] - Baily, l. / 1979 / Egypt bee virus and Australian isolates of kashmir bee virus / J. Gen. Virol 43 : 641 ~ 647
4. [학술지(정기간행물)] - Chen, Y.P. / 2004 / Molecular evidence for transmission of Kashmiri bee virus in honey bee colonis by ectoparasitic mite, Varroa jacobsoni Oud, under laboratory conditions / Apidologie 35 : 441 ~ 448
5. [학술지(정기간행물)] - Grabernsteiner, E. / 2001 / Sacbrood Virus of the honeybee (Apis mellifera): Rapid Identification and phylogenetic Analysis Using Revers Transcription PCR / Clinical and Diagnostic Laboratory Immunology : 83 ~ 104
2. [단행본] - Baily, L. / 1991 / Honey bee pathology / Academic Press
3. [학술지(정기간행물)] - Baily, l. / 1979 / Egypt bee virus and Australian isolates of kashmir bee virus / J. Gen. Virol 43 : 641 ~ 647
4. [학술지(정기간행물)] - Chen, Y.P. / 2004 / Molecular evidence for transmission of Kashmiri bee virus in honey bee colonis by ectoparasitic mite, Varroa jacobsoni Oud, under laboratory conditions / Apidologie 35 : 441 ~ 448
5. [학술지(정기간행물)] - Grabernsteiner, E. / 2001 / Sacbrood Virus of the honeybee (Apis mellifera): Rapid Identification and phylogenetic Analysis Using Revers Transcription PCR / Clinical and Diagnostic Laboratory Immunology : 83 ~ 104
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