Multiple pulses improve electroporation efficiency in Agrobacterium tumefaciens

Electroporation entails brief, high intensity pulse to create transient pores in the cell membrane to facilitate the entry of exogenous macromolecules, which may otherwise be excluded. Removal of the external field leads to the resealing of the membrane electropores permitting the survival of the el...

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Autores principales:	Mahmood,Tariq, Zar,Tamkina, Saqlan Naqvi,S.M
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2008
Materias:	Agrobacterium electroporation pulse number transformation efficiency
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100015
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spelling	oai:scielo:S0717-345820080001000152008-10-28Multiple pulses improve electroporation efficiency in Agrobacterium tumefaciensMahmood,TariqZar,TamkinaSaqlan Naqvi,S.M Agrobacterium electroporation pulse number transformation efficiency Electroporation entails brief, high intensity pulse to create transient pores in the cell membrane to facilitate the entry of exogenous macromolecules, which may otherwise be excluded. Removal of the external field leads to the resealing of the membrane electropores permitting the survival of the electrically stimulated recipient cells. Using this technique foreign deoxyribonucleic acid (DNA) has been successfully introduced into many cell types both from prokaryotes and eukaryotes. Increase in pulse voltage and length beyond a critical limit has been reported to decrease transformation efficiency, hence in this study we have investigated another strategy i.e. increase in the number of pulses at constant high voltage and pulse duration. Commonly used Agrobacterium strains LBA4404 and EHA101 and binary vector pCAMBIA1301 were used. Transformants were selected on a combination of hygromycin and kanamycin, and confirmed by polymerase chain reaction (PCR) and restriction analysis. Increase in the number of pulses was found to show a significant and linear increase in transformation efficiency.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.11 n.1 20082008-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100015en10.4067/S0717-34582008000100015
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Agrobacterium electroporation pulse number transformation efficiency
spellingShingle	Agrobacterium electroporation pulse number transformation efficiency Mahmood,Tariq Zar,Tamkina Saqlan Naqvi,S.M Multiple pulses improve electroporation efficiency in Agrobacterium tumefaciens
description	Electroporation entails brief, high intensity pulse to create transient pores in the cell membrane to facilitate the entry of exogenous macromolecules, which may otherwise be excluded. Removal of the external field leads to the resealing of the membrane electropores permitting the survival of the electrically stimulated recipient cells. Using this technique foreign deoxyribonucleic acid (DNA) has been successfully introduced into many cell types both from prokaryotes and eukaryotes. Increase in pulse voltage and length beyond a critical limit has been reported to decrease transformation efficiency, hence in this study we have investigated another strategy i.e. increase in the number of pulses at constant high voltage and pulse duration. Commonly used Agrobacterium strains LBA4404 and EHA101 and binary vector pCAMBIA1301 were used. Transformants were selected on a combination of hygromycin and kanamycin, and confirmed by polymerase chain reaction (PCR) and restriction analysis. Increase in the number of pulses was found to show a significant and linear increase in transformation efficiency.
author	Mahmood,Tariq Zar,Tamkina Saqlan Naqvi,S.M
author_facet	Mahmood,Tariq Zar,Tamkina Saqlan Naqvi,S.M
author_sort	Mahmood,Tariq
title	Multiple pulses improve electroporation efficiency in Agrobacterium tumefaciens
title_short	Multiple pulses improve electroporation efficiency in Agrobacterium tumefaciens
title_full	Multiple pulses improve electroporation efficiency in Agrobacterium tumefaciens
title_fullStr	Multiple pulses improve electroporation efficiency in Agrobacterium tumefaciens
title_full_unstemmed	Multiple pulses improve electroporation efficiency in Agrobacterium tumefaciens
title_sort	multiple pulses improve electroporation efficiency in agrobacterium tumefaciens
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2008
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100015
work_keys_str_mv	AT mahmoodtariq multiplepulsesimproveelectroporationefficiencyinagrobacteriumtumefaciens AT zartamkina multiplepulsesimproveelectroporationefficiencyinagrobacteriumtumefaciens AT saqlannaqvism multiplepulsesimproveelectroporationefficiencyinagrobacteriumtumefaciens
_version_	1718441781112078336

Multiple pulses improve electroporation efficiency in Agrobacterium tumefaciens

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