A simple negative selection method to identify adenovirus recombinants using colony PCR

Background: The AdEasy system is a fast-track system for generating recombinant adenoviruses using the efficient homologous recombination machinery between shuttle and adenovirus backbone plasmids in Escherichia coli BJ5183 cells. The key step is homologous recombination in BJ5183 cells, which is dr...

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Autores principales:	Zhao,Yongliang, Dong,Zongming, Li,Tengfei, Huang,Gang
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2014
Materias:	Adenovirus AdEasy Colony PCR Homologous recombination RecA
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000100008
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spelling	oai:scielo:S0717-345820140001000082014-04-07A simple negative selection method to identify adenovirus recombinants using colony PCRZhao,YongliangDong,ZongmingLi,TengfeiHuang,Gang Adenovirus AdEasy Colony PCR Homologous recombination RecA Background: The AdEasy system is a fast-track system for generating recombinant adenoviruses using the efficient homologous recombination machinery between shuttle and adenovirus backbone plasmids in Escherichia coli BJ5183 cells. The key step is homologous recombination in BJ5183 cells, which is driven by RecA activity. However, culture time is stringently limited to reduce the damage to recombinant plasmids by RecA activity. Therefore, rapid identification of recombinant adenoviruses within the limited time-period is critical. Results: We developed a simple negative selection method to identify recombinant adenoviruses using colony PCR, which improves the efficiency of adenovirus recombination screening and packaging. Conclusions: The negative selection method to identify AdEasy adenovirus recombinants by colony PCR can identify the recombined colony within a short time-period, and maximally avoid damage to the recombinant plasmid by limiting recombination time, resulting in improved adenovirus packaging.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.17 n.1 20142014-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000100008en
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Adenovirus AdEasy Colony PCR Homologous recombination RecA
spellingShingle	Adenovirus AdEasy Colony PCR Homologous recombination RecA Zhao,Yongliang Dong,Zongming Li,Tengfei Huang,Gang A simple negative selection method to identify adenovirus recombinants using colony PCR
description	Background: The AdEasy system is a fast-track system for generating recombinant adenoviruses using the efficient homologous recombination machinery between shuttle and adenovirus backbone plasmids in Escherichia coli BJ5183 cells. The key step is homologous recombination in BJ5183 cells, which is driven by RecA activity. However, culture time is stringently limited to reduce the damage to recombinant plasmids by RecA activity. Therefore, rapid identification of recombinant adenoviruses within the limited time-period is critical. Results: We developed a simple negative selection method to identify recombinant adenoviruses using colony PCR, which improves the efficiency of adenovirus recombination screening and packaging. Conclusions: The negative selection method to identify AdEasy adenovirus recombinants by colony PCR can identify the recombined colony within a short time-period, and maximally avoid damage to the recombinant plasmid by limiting recombination time, resulting in improved adenovirus packaging.
author	Zhao,Yongliang Dong,Zongming Li,Tengfei Huang,Gang
author_facet	Zhao,Yongliang Dong,Zongming Li,Tengfei Huang,Gang
author_sort	Zhao,Yongliang
title	A simple negative selection method to identify adenovirus recombinants using colony PCR
title_short	A simple negative selection method to identify adenovirus recombinants using colony PCR
title_full	A simple negative selection method to identify adenovirus recombinants using colony PCR
title_fullStr	A simple negative selection method to identify adenovirus recombinants using colony PCR
title_full_unstemmed	A simple negative selection method to identify adenovirus recombinants using colony PCR
title_sort	simple negative selection method to identify adenovirus recombinants using colony pcr
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2014
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582014000100008
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A simple negative selection method to identify adenovirus recombinants using colony PCR

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