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Characterization of the duck enteritis virus UL55 protein

BACKGROUND: Characteration of the newly identified duck enteritis virus UL55 gene product has not been reported yet. Knowledge of the protein UL55 can provide useful insights about its function. RESULTS: The newly identified duck enteritis virus UL55 gene was about 561 bp, it was amplified and diges...

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Autores principales: Wu, Ying, Cheng, Anchun, Wang, Mingshu, Zhang, Shunchuan, Zhu, Dekang, Jia, Renyong, Luo, Qihui, Chen, Zhengli, Chen, Xiaoyue
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3117846/
https://www.ncbi.nlm.nih.gov/pubmed/21609474
http://dx.doi.org/10.1186/1743-422X-8-256
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author Wu, Ying
Cheng, Anchun
Wang, Mingshu
Zhang, Shunchuan
Zhu, Dekang
Jia, Renyong
Luo, Qihui
Chen, Zhengli
Chen, Xiaoyue
author_facet Wu, Ying
Cheng, Anchun
Wang, Mingshu
Zhang, Shunchuan
Zhu, Dekang
Jia, Renyong
Luo, Qihui
Chen, Zhengli
Chen, Xiaoyue
author_sort Wu, Ying
collection PubMed
description BACKGROUND: Characteration of the newly identified duck enteritis virus UL55 gene product has not been reported yet. Knowledge of the protein UL55 can provide useful insights about its function. RESULTS: The newly identified duck enteritis virus UL55 gene was about 561 bp, it was amplified and digested for construction of a recombinant plasmid pET32a(+)/UL55 for expression in Escherichia coli. SDS-PAGE analysis revealed the recombinant protein UL55(pUL55) was overexpressed in Escherichia coli BL21 host cells after induction by 0.2 mM IPTG at 37°C for 4 h and aggregated as inclusion bodies. The denatured protein about 40 KDa named pUL55 was purified by washing five times, and used to immune rabbits for preparation of polyclonal antibody. The prepared polyclonal antibody against pUL55 was detected and determined by Agar immundiffusion and Neutralization test. The results of Wstern blotting assay and intracellular analysis revealed that pUL55 was expressed most abundantly during the late phase of replication and mainly distributed in cytoplasm in duck enteritis virus infected cells. CONCLUSIONS: In this study, the duck enteritis virus UL55 protein was successfully expressed in prokaryotic expression system. Besides, we have prepared the polyclonal antibody against recombinant prtein UL55, and characterized some properties of the duck enteritis virus UL55 protein for the first time. The research will be useful for further functional analysis of this gene.
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spelling pubmed-31178462011-06-18 Characterization of the duck enteritis virus UL55 protein Wu, Ying Cheng, Anchun Wang, Mingshu Zhang, Shunchuan Zhu, Dekang Jia, Renyong Luo, Qihui Chen, Zhengli Chen, Xiaoyue Virol J Research BACKGROUND: Characteration of the newly identified duck enteritis virus UL55 gene product has not been reported yet. Knowledge of the protein UL55 can provide useful insights about its function. RESULTS: The newly identified duck enteritis virus UL55 gene was about 561 bp, it was amplified and digested for construction of a recombinant plasmid pET32a(+)/UL55 for expression in Escherichia coli. SDS-PAGE analysis revealed the recombinant protein UL55(pUL55) was overexpressed in Escherichia coli BL21 host cells after induction by 0.2 mM IPTG at 37°C for 4 h and aggregated as inclusion bodies. The denatured protein about 40 KDa named pUL55 was purified by washing five times, and used to immune rabbits for preparation of polyclonal antibody. The prepared polyclonal antibody against pUL55 was detected and determined by Agar immundiffusion and Neutralization test. The results of Wstern blotting assay and intracellular analysis revealed that pUL55 was expressed most abundantly during the late phase of replication and mainly distributed in cytoplasm in duck enteritis virus infected cells. CONCLUSIONS: In this study, the duck enteritis virus UL55 protein was successfully expressed in prokaryotic expression system. Besides, we have prepared the polyclonal antibody against recombinant prtein UL55, and characterized some properties of the duck enteritis virus UL55 protein for the first time. The research will be useful for further functional analysis of this gene. BioMed Central 2011-05-24 /pmc/articles/PMC3117846/ /pubmed/21609474 http://dx.doi.org/10.1186/1743-422X-8-256 Text en Copyright ©2011 Wu et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Wu, Ying
Cheng, Anchun
Wang, Mingshu
Zhang, Shunchuan
Zhu, Dekang
Jia, Renyong
Luo, Qihui
Chen, Zhengli
Chen, Xiaoyue
Characterization of the duck enteritis virus UL55 protein
title Characterization of the duck enteritis virus UL55 protein
title_full Characterization of the duck enteritis virus UL55 protein
title_fullStr Characterization of the duck enteritis virus UL55 protein
title_full_unstemmed Characterization of the duck enteritis virus UL55 protein
title_short Characterization of the duck enteritis virus UL55 protein
title_sort characterization of the duck enteritis virus ul55 protein
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3117846/
https://www.ncbi.nlm.nih.gov/pubmed/21609474
http://dx.doi.org/10.1186/1743-422X-8-256
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