Location and cloning of the ketal pyruvate transferase gene of Xanthomonas campestris

Genes required for xanthan polysaccharide synthesis (xps) are clustered in a DNA region of 13.5 kb in the chromosome of Xanthomonas campestris. Plasmid pCHC3 containing a 12.4-kb insert of xps genes has been suggested to include a gene involved in the pyruvylation of xanthan gum (N. E. Harding, J. M...

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Autores principales: Petroni, Eberto Alejandro, Ielpi, Luis
Publicado: 1991
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Acceso en línea:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219193_v173_n23_p7519_Marzocca
http://hdl.handle.net/20.500.12110/paper_00219193_v173_n23_p7519_Marzocca
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spelling paper:paper_00219193_v173_n23_p7519_Marzocca2023-06-08T14:43:09Z Location and cloning of the ketal pyruvate transferase gene of Xanthomonas campestris Petroni, Eberto Alejandro Ielpi, Luis ketal pyruvate transferase unclassified drug xanthan article chromosome gene mapping molecular cloning nonhuman plasmid priority journal Xanthomonas Xanthomonas campestris Genes required for xanthan polysaccharide synthesis (xps) are clustered in a DNA region of 13.5 kb in the chromosome of Xanthomonas campestris. Plasmid pCHC3 containing a 12.4-kb insert of xps genes has been suggested to include a gene involved in the pyruvylation of xanthan gum (N. E. Harding, J. M. Cleary, D. K. Cabanas, I. G. Rosen, and K. S. Kang, J. Bacteriol. 169:2854-2861, 1987). An essential step toward understanding the biosynthesis of xanthan gum and to enable genetic manipulation of xanthan structure is the determination of the biochemical function encoded by the xps genes. On the basis of biochemical characterization of an X. campestris mutant which produces pyruvate-free xanthan gum, complementation studies, and heterologous expression, we have identified the gene coding for the ketal pyruvate transferase (kpt) enzyme. This gene was located on a 1.4-kb BamHI fragment of pCHC3 and cloned in the broad-host-range cloning vector pRK404. An X. campestris kpt mutant was constructed by mini-Mu(Tet(r)) mutagenesis of the cloned gene and then by recombination of the mutation into the chromosome of the wild-type strain. Fil:Petroni, E.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Ielpi, L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1991 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219193_v173_n23_p7519_Marzocca http://hdl.handle.net/20.500.12110/paper_00219193_v173_n23_p7519_Marzocca
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic ketal pyruvate transferase
unclassified drug
xanthan
article
chromosome
gene mapping
molecular cloning
nonhuman
plasmid
priority journal
Xanthomonas
Xanthomonas campestris
spellingShingle ketal pyruvate transferase
unclassified drug
xanthan
article
chromosome
gene mapping
molecular cloning
nonhuman
plasmid
priority journal
Xanthomonas
Xanthomonas campestris
Petroni, Eberto Alejandro
Ielpi, Luis
Location and cloning of the ketal pyruvate transferase gene of Xanthomonas campestris
topic_facet ketal pyruvate transferase
unclassified drug
xanthan
article
chromosome
gene mapping
molecular cloning
nonhuman
plasmid
priority journal
Xanthomonas
Xanthomonas campestris
description Genes required for xanthan polysaccharide synthesis (xps) are clustered in a DNA region of 13.5 kb in the chromosome of Xanthomonas campestris. Plasmid pCHC3 containing a 12.4-kb insert of xps genes has been suggested to include a gene involved in the pyruvylation of xanthan gum (N. E. Harding, J. M. Cleary, D. K. Cabanas, I. G. Rosen, and K. S. Kang, J. Bacteriol. 169:2854-2861, 1987). An essential step toward understanding the biosynthesis of xanthan gum and to enable genetic manipulation of xanthan structure is the determination of the biochemical function encoded by the xps genes. On the basis of biochemical characterization of an X. campestris mutant which produces pyruvate-free xanthan gum, complementation studies, and heterologous expression, we have identified the gene coding for the ketal pyruvate transferase (kpt) enzyme. This gene was located on a 1.4-kb BamHI fragment of pCHC3 and cloned in the broad-host-range cloning vector pRK404. An X. campestris kpt mutant was constructed by mini-Mu(Tet(r)) mutagenesis of the cloned gene and then by recombination of the mutation into the chromosome of the wild-type strain.
author Petroni, Eberto Alejandro
Ielpi, Luis
author_facet Petroni, Eberto Alejandro
Ielpi, Luis
author_sort Petroni, Eberto Alejandro
title Location and cloning of the ketal pyruvate transferase gene of Xanthomonas campestris
title_short Location and cloning of the ketal pyruvate transferase gene of Xanthomonas campestris
title_full Location and cloning of the ketal pyruvate transferase gene of Xanthomonas campestris
title_fullStr Location and cloning of the ketal pyruvate transferase gene of Xanthomonas campestris
title_full_unstemmed Location and cloning of the ketal pyruvate transferase gene of Xanthomonas campestris
title_sort location and cloning of the ketal pyruvate transferase gene of xanthomonas campestris
publishDate 1991
url https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219193_v173_n23_p7519_Marzocca
http://hdl.handle.net/20.500.12110/paper_00219193_v173_n23_p7519_Marzocca
work_keys_str_mv AT petroniebertoalejandro locationandcloningoftheketalpyruvatetransferasegeneofxanthomonascampestris
AT ielpiluis locationandcloningoftheketalpyruvatetransferasegeneofxanthomonascampestris
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