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Characterization of a small cryptic plasmid from endophytic Pantoea agglomerans and its use in the construction of an expression vector

A circular cryptic plasmid named pPAGA (2,734 bp) was isolated from Pantoea agglomerans strain EGE6 (an endophytic bacterial isolate from eucalyptus). Sequence analysis revealed that the plasmid has a G+C content of 51% and contains four potential ORFs, 238(A), 250(B), 131(C), and 129(D) amino acids...

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Autores principales: de Lima Procópio, Rudi Emerson, Araújo, Welington Luiz, Andreote, Fernando Dini, Azevedo, João Lúcio
Formato: Texto
Lenguaje:English
Publicado: Sociedade Brasileira de Genética 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3085353/
https://www.ncbi.nlm.nih.gov/pubmed/21637551
http://dx.doi.org/10.1590/S1415-47572010005000096
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author de Lima Procópio, Rudi Emerson
Araújo, Welington Luiz
Andreote, Fernando Dini
Azevedo, João Lúcio
author_facet de Lima Procópio, Rudi Emerson
Araújo, Welington Luiz
Andreote, Fernando Dini
Azevedo, João Lúcio
author_sort de Lima Procópio, Rudi Emerson
collection PubMed
description A circular cryptic plasmid named pPAGA (2,734 bp) was isolated from Pantoea agglomerans strain EGE6 (an endophytic bacterial isolate from eucalyptus). Sequence analysis revealed that the plasmid has a G+C content of 51% and contains four potential ORFs, 238(A), 250(B), 131(C), and 129(D) amino acids in length without homology to known proteins. The shuttle vector pLGM1 was constructed by combining the pPAGA plasmid with pGFPmut3.0 (which harbors a gene encoding green fluorescent protein, GFP), and the resulting construct was used to over-express GFP in E. coli and P. agglomerans cells. GFP production was used to monitor the colonization of strain EGE6gfp in various plant tissues by fluorescence microscopy. Analysis of EGE6gfp colonization showed that 14 days after inoculation, the strain occupied the inner tissue of Eucalyptus grandis roots, preferentially colonizing the xylem vessels of the host plants.
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spelling pubmed-30853532011-06-02 Characterization of a small cryptic plasmid from endophytic Pantoea agglomerans and its use in the construction of an expression vector de Lima Procópio, Rudi Emerson Araújo, Welington Luiz Andreote, Fernando Dini Azevedo, João Lúcio Genet Mol Biol Genetics of Microorganisms A circular cryptic plasmid named pPAGA (2,734 bp) was isolated from Pantoea agglomerans strain EGE6 (an endophytic bacterial isolate from eucalyptus). Sequence analysis revealed that the plasmid has a G+C content of 51% and contains four potential ORFs, 238(A), 250(B), 131(C), and 129(D) amino acids in length without homology to known proteins. The shuttle vector pLGM1 was constructed by combining the pPAGA plasmid with pGFPmut3.0 (which harbors a gene encoding green fluorescent protein, GFP), and the resulting construct was used to over-express GFP in E. coli and P. agglomerans cells. GFP production was used to monitor the colonization of strain EGE6gfp in various plant tissues by fluorescence microscopy. Analysis of EGE6gfp colonization showed that 14 days after inoculation, the strain occupied the inner tissue of Eucalyptus grandis roots, preferentially colonizing the xylem vessels of the host plants. Sociedade Brasileira de Genética 2011 2011-03-01 /pmc/articles/PMC3085353/ /pubmed/21637551 http://dx.doi.org/10.1590/S1415-47572010005000096 Text en Copyright © 2011, Sociedade Brasileira de Genética. License information: This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Genetics of Microorganisms
de Lima Procópio, Rudi Emerson
Araújo, Welington Luiz
Andreote, Fernando Dini
Azevedo, João Lúcio
Characterization of a small cryptic plasmid from endophytic Pantoea agglomerans and its use in the construction of an expression vector
title Characterization of a small cryptic plasmid from endophytic Pantoea agglomerans and its use in the construction of an expression vector
title_full Characterization of a small cryptic plasmid from endophytic Pantoea agglomerans and its use in the construction of an expression vector
title_fullStr Characterization of a small cryptic plasmid from endophytic Pantoea agglomerans and its use in the construction of an expression vector
title_full_unstemmed Characterization of a small cryptic plasmid from endophytic Pantoea agglomerans and its use in the construction of an expression vector
title_short Characterization of a small cryptic plasmid from endophytic Pantoea agglomerans and its use in the construction of an expression vector
title_sort characterization of a small cryptic plasmid from endophytic pantoea agglomerans and its use in the construction of an expression vector
topic Genetics of Microorganisms
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3085353/
https://www.ncbi.nlm.nih.gov/pubmed/21637551
http://dx.doi.org/10.1590/S1415-47572010005000096
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