A simple method for in vivo expression studies of Xanthomonas axonopodis pv. citri
Identifieur interne : 002B78 ( Main/Exploration ); précédent : 002B77; suivant : 002B79A simple method for in vivo expression studies of Xanthomonas axonopodis pv. citri
Auteurs : Angela Mehta [Brésil] ; Yoko B. Rosato [Brésil]Source :
- Current microbiology : (Print) [ 0343-8651 ] ; 2003.
Descripteurs français
- Pascal (Inist)
English descriptors
- KwdEn :
- 16S-RNA, Bacterial Proteins (analysis), Bacterial Proteins (genetics), Bacterial Proteins (isolation & purification), Bacteriosis, Citrus sinensis, Citrus sinensis (microbiology), DNA-Binding Proteins (genetics), Electrophoresis, Gel, Two-Dimensional (methods), Fingerprint method, Gene Expression, Isolate, Messenger RNA, Plant Leaves (microbiology), Plant pathogen, Protein, Proteome (analysis), Proteome (isolation & purification), RNA, Bacterial (analysis), RNA, Bacterial (isolation & purification), RNA, Ribosomal (analysis), RNA, Ribosomal (isolation & purification), Reverse Transcriptase Polymerase Chain Reaction (methods), Reverse transcription polymerase chain reaction, Ribosomal RNA, Transcription Factors (genetics), Transcription, Genetic, Xanthomonas (chemistry), Xanthomonas (genetics), Xanthomonas (isolation & purification), Xanthomonas axonopodis.
- MESH :
- chemical , analysis : Bacterial Proteins, Proteome, RNA, Bacterial, RNA, Ribosomal.
- chemical , genetics : Bacterial Proteins, DNA-Binding Proteins, Transcription Factors.
- chemical , isolation & purification : Bacterial Proteins, Proteome, RNA, Bacterial, RNA, Ribosomal.
- chemistry : Xanthomonas.
- genetics : Xanthomonas.
- isolation & purification : Xanthomonas.
- methods : Electrophoresis, Gel, Two-Dimensional, Reverse Transcriptase Polymerase Chain Reaction.
- microbiology : Citrus sinensis, Plant Leaves.
- Gene Expression, Transcription, Genetic.
Abstract
A major problem in studying bacterial plant pathogens is obtaining the microorganism directly from the plant tissue to perform in vivo expression (protein or mRNA) analyses. Here we report an easy and fast protocol to isolate Xanthomonas axonopodis pv. citri directly from the host plant, in sufficient amounts to perform protein fingerprinting by 2-D gel electrophoresis as well as RNA expression assays. The protein profile obtained was very similar to that of X. axonopodis pv. citri grown in the presence of a leaf extract of Citrus sinensis; however, some differential proteins expressed in vivo were observed. Total RNA extraction revealed typical 16S and 23S bands in the agarose gel, and RT-PCR reactions using primers specific for genes of the bacterium confirmed the quality of the RNA preparation. Also, RT-PCR reactions using plant ribosomal primers were employed, and no amplification product was obtained, indicating that plant RNA is not present in the bacterium RNA sample.
Affiliations:
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Rosato</name><affiliation wicri:level="2"><inist:fA14 i1="02"><s1>CBMEG-UNICAMP, Caixa Postal 6010</s1><s2>CEP 13083-970, Campinas, SP</s2><s3>BRA</s3><sZ>2 aut.</sZ></inist:fA14><country>Brésil</country><placeName><region type="state">État de São Paulo</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">04-0314193</idno><date when="2003">2003</date><idno type="stanalyst">PASCAL 04-0314193 INIST</idno><idno type="RBID">Pascal:04-0314193</idno><idno type="wicri:Area/PascalFrancis/Corpus">000696</idno><idno type="wicri:Area/PascalFrancis/Curation">000308</idno><idno type="wicri:Area/PascalFrancis/Checkpoint">000761</idno><idno type="wicri:doubleKey">0343-8651:2003:Mehta A:a:simple:method</idno><idno type="wicri:Area/Main/Merge">002D47</idno><idno type="wicri:source">PubMed</idno><idno type="RBID">pubmed:14669917</idno><idno type="wicri:Area/PubMed/Corpus">000E20</idno><idno type="wicri:Area/PubMed/Curation">000E20</idno><idno type="wicri:Area/PubMed/Checkpoint">000E20</idno><idno type="wicri:Area/Ncbi/Merge">000240</idno><idno type="wicri:Area/Ncbi/Curation">000240</idno><idno type="wicri:Area/Ncbi/Checkpoint">000240</idno><idno type="wicri:doubleKey">0343-8651:2003:Mehta A:a:simple:method</idno><idno type="wicri:Area/Main/Merge">002C29</idno><idno type="wicri:Area/Main/Curation">002B78</idno><idno type="wicri:Area/Main/Exploration">002B78</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">A simple method for in vivo expression studies of Xanthomonas axonopodis pv. citri</title><author><name sortKey="Mehta, Angela" sort="Mehta, Angela" uniqKey="Mehta A" first="Angela" last="Mehta">Angela Mehta</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>EMBRAPA-CENARGEN, Parque Estação Biológica Final Av. W/5 Norte</s1><s2>CEP 70770-900, Brasilia, DF</s2><s3>BRA</s3><sZ>1 aut.</sZ></inist:fA14><country>Brésil</country><placeName><region type="state">District fédéral (Brésil)</region></placeName></affiliation></author><author><name sortKey="Rosato, Yoko B" sort="Rosato, Yoko B" uniqKey="Rosato Y" first="Yoko B." last="Rosato">Yoko B. Rosato</name><affiliation wicri:level="2"><inist:fA14 i1="02"><s1>CBMEG-UNICAMP, Caixa Postal 6010</s1><s2>CEP 13083-970, Campinas, SP</s2><s3>BRA</s3><sZ>2 aut.</sZ></inist:fA14><country>Brésil</country><placeName><region type="state">État de São Paulo</region></placeName></affiliation></author></analytic><series><title level="j" type="main">Current microbiology : (Print)</title><title level="j" type="abbreviated">Cur. microbiol. : (Print)</title><idno type="ISSN">0343-8651</idno><imprint><date when="2003">2003</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Current microbiology : (Print)</title><title level="j" type="abbreviated">Cur. microbiol. : (Print)</title><idno type="ISSN">0343-8651</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>16S-RNA</term><term>Bacterial Proteins (analysis)</term><term>Bacterial Proteins (genetics)</term><term>Bacterial Proteins (isolation & purification)</term><term>Bacteriosis</term><term>Citrus sinensis</term><term>Citrus sinensis (microbiology)</term><term>DNA-Binding Proteins (genetics)</term><term>Electrophoresis, Gel, Two-Dimensional (methods)</term><term>Fingerprint method</term><term>Gene Expression</term><term>Isolate</term><term>Messenger RNA</term><term>Plant Leaves (microbiology)</term><term>Plant pathogen</term><term>Protein</term><term>Proteome (analysis)</term><term>Proteome (isolation & purification)</term><term>RNA, Bacterial (analysis)</term><term>RNA, Bacterial (isolation & purification)</term><term>RNA, Ribosomal (analysis)</term><term>RNA, Ribosomal (isolation & purification)</term><term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term><term>Reverse transcription polymerase chain reaction</term><term>Ribosomal RNA</term><term>Transcription Factors (genetics)</term><term>Transcription, Genetic</term><term>Xanthomonas (chemistry)</term><term>Xanthomonas (genetics)</term><term>Xanthomonas (isolation & purification)</term><term>Xanthomonas axonopodis</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Bacterial Proteins</term><term>Proteome</term><term>RNA, Bacterial</term><term>RNA, Ribosomal</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Bacterial Proteins</term><term>DNA-Binding Proteins</term><term>Transcription Factors</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Bacterial Proteins</term><term>Proteome</term><term>RNA, Bacterial</term><term>RNA, Ribosomal</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Xanthomonas</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Xanthomonas</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Xanthomonas</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Electrophoresis, Gel, Two-Dimensional</term><term>Reverse Transcriptase Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Citrus sinensis</term><term>Plant Leaves</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Expression</term><term>Transcription, Genetic</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Xanthomonas axonopodis</term><term>Citrus sinensis</term><term>Méthode fingerprint</term><term>Phytopathogène</term><term>Protéine</term><term>RNA messager</term><term>Isolat</term><term>RNA 16S</term><term>RNA ribosomique</term><term>Réaction chaîne polymérase RT</term><term>Bactériose</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A major problem in studying bacterial plant pathogens is obtaining the microorganism directly from the plant tissue to perform in vivo expression (protein or mRNA) analyses. Here we report an easy and fast protocol to isolate Xanthomonas axonopodis pv. citri directly from the host plant, in sufficient amounts to perform protein fingerprinting by 2-D gel electrophoresis as well as RNA expression assays. The protein profile obtained was very similar to that of X. axonopodis pv. citri grown in the presence of a leaf extract of Citrus sinensis; however, some differential proteins expressed in vivo were observed. Total RNA extraction revealed typical 16S and 23S bands in the agarose gel, and RT-PCR reactions using primers specific for genes of the bacterium confirmed the quality of the RNA preparation. Also, RT-PCR reactions using plant ribosomal primers were employed, and no amplification product was obtained, indicating that plant RNA is not present in the bacterium RNA sample.</div></front></TEI><affiliations><list><country><li>Brésil</li></country><region><li>District fédéral (Brésil)</li><li>État de São Paulo</li></region></list><tree><country name="Brésil"><region name="District fédéral (Brésil)"><name sortKey="Mehta, Angela" sort="Mehta, Angela" uniqKey="Mehta A" first="Angela" last="Mehta">Angela Mehta</name></region><name sortKey="Rosato, Yoko B" sort="Rosato, Yoko B" uniqKey="Rosato Y" first="Yoko B." last="Rosato">Yoko B. Rosato</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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