Design and expression of a retro doublet of cecropin with enhanced activity

Novel doublet molecules of cecropin A from Drosophila melanogaster were designed and constructed combining the regular (CECdir) with the inverted (CECret) coding sequence of the standard CEC A1 gene resulting in the following configurations: CECdir-CECret and CECret-CECdir. These two recombinant mol...

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Autores principales:	Díaz,Mauricio, Arenas,Gloria, Marshall,Sergio H
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2008
Materias:	antimicrobial peptides Escherichia coli expression
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000200006
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spelling	oai:scielo:S0717-345820080002000062009-01-21Design and expression of a retro doublet of cecropin with enhanced activityDíaz,MauricioArenas,GloriaMarshall,Sergio H antimicrobial peptides Escherichia coli expression Novel doublet molecules of cecropin A from Drosophila melanogaster were designed and constructed combining the regular (CECdir) with the inverted (CECret) coding sequence of the standard CEC A1 gene resulting in the following configurations: CECdir-CECret and CECret-CECdir. These two recombinant molecules were generated using a three-primer driven PCR reaction yielding composite single functional aminoacidic molecules with the coding sequences of CECdir linked in frame with the coding sequence of CECret and vice versa. In order to obtain these constructions, a retropeptide DNA-coding sequence was chemically synthesized to match the expected polarity of the newly generated CECret sequence. Both doublet antimicrobial peptides (drAMPs) were cloned in the T7 promoter driven expression plasmid pET27b+ and expressed in E. coli BL21 without any fusion protein. Only the former recombinant peptide was expressed and purified from cell extracts and its specific activity against two different bacteria showed to be higher than those displayed by their monomer parental counterparts.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.11 n.2 20082008-04-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000200006en10.4067/S0717-34582008000200006
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	antimicrobial peptides Escherichia coli expression
spellingShingle	antimicrobial peptides Escherichia coli expression Díaz,Mauricio Arenas,Gloria Marshall,Sergio H Design and expression of a retro doublet of cecropin with enhanced activity
description	Novel doublet molecules of cecropin A from Drosophila melanogaster were designed and constructed combining the regular (CECdir) with the inverted (CECret) coding sequence of the standard CEC A1 gene resulting in the following configurations: CECdir-CECret and CECret-CECdir. These two recombinant molecules were generated using a three-primer driven PCR reaction yielding composite single functional aminoacidic molecules with the coding sequences of CECdir linked in frame with the coding sequence of CECret and vice versa. In order to obtain these constructions, a retropeptide DNA-coding sequence was chemically synthesized to match the expected polarity of the newly generated CECret sequence. Both doublet antimicrobial peptides (drAMPs) were cloned in the T7 promoter driven expression plasmid pET27b+ and expressed in E. coli BL21 without any fusion protein. Only the former recombinant peptide was expressed and purified from cell extracts and its specific activity against two different bacteria showed to be higher than those displayed by their monomer parental counterparts.
author	Díaz,Mauricio Arenas,Gloria Marshall,Sergio H
author_facet	Díaz,Mauricio Arenas,Gloria Marshall,Sergio H
author_sort	Díaz,Mauricio
title	Design and expression of a retro doublet of cecropin with enhanced activity
title_short	Design and expression of a retro doublet of cecropin with enhanced activity
title_full	Design and expression of a retro doublet of cecropin with enhanced activity
title_fullStr	Design and expression of a retro doublet of cecropin with enhanced activity
title_full_unstemmed	Design and expression of a retro doublet of cecropin with enhanced activity
title_sort	design and expression of a retro doublet of cecropin with enhanced activity
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2008
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000200006
work_keys_str_mv	AT diazmauricio designandexpressionofaretrodoubletofcecropinwithenhancedactivity AT arenasgloria designandexpressionofaretrodoubletofcecropinwithenhancedactivity AT marshallsergioh designandexpressionofaretrodoubletofcecropinwithenhancedactivity
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Design and expression of a retro doublet of cecropin with enhanced activity

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