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The spliceosomal intron of the rolA gene of Agrobacterium rhizogenes is a prokaryotic promoter
Author(s) -
Pandolfini T.,
Storlazzi A.,
Calabria E.,
Defez R.,
Spena A.
Publication year - 2000
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1046/j.1365-2958.2000.01810.x
Subject(s) - biology , agrobacterium , gene , intron , reporter gene , genetics , plasmid , microbiology and biotechnology , gene expression , promoter , gus reporter system , dna , chimeric gene , transformation (genetics)
Agrobacterium rhizogenes transfers DNA (T‐DNA) from its Ri plasmid to plant cells. All T‐DNA genes are expressed in plant cells. The rolA gene is the only T‐DNA gene that contains an intron in the untranslated leader region of its mRNA. This paper shows that (i) the rolA gene is also transcribed in bacteria; (ii) the 85 bp corresponding to the spliceosomal intron drives prokaryotic gene expression in agrobacteria, in free‐living rhizobia and in bacteroids within root nodules; and (iii) promoter activity is abolished by the deletion of 63 bp from its 5′ end and is reduced by mutations changing its sequence near the putative −10 region. The expression pattern of a chimeric reporter gene shows that, in free‐living bacteria, gene expression takes place during the exponential phase of growth and increases at the onset of the stationary phase. Within root nodules, reporter gene expression occurs in the invasion, nitrogen fixing and senescent zones.