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MONITORED ENVIRONMENT RELEASE OF GENOMICALLY TAGGED PHYLLOPLANE AND RIZOSPHERE BACTERIA

Objetivo


P. aureofaciens SBW25, a plasmid free, nonpathogen, was the dominant species (<30%) of bacteria found in the phylloplane of sugar beet. SBW25 was chromosomally marked with pMOB::TN5XYLE, to confirm the suitability of SWB25, the markers (Kmr, XylE) and to develop the plant model. Recombinants' free of any mobilisable elements or plasmid sequence were made; SBW25 marked by the insertion of a Kmr gene (From Tn903) into a known, nonfunctional chromosomal site to create safe, environmentally acceptable genetically modified organisms (GMOs) as competitively fit as the wild type. Recombinants were indistinguishable from wild type in mono or mixed cultures, with no loss of phenotype recorded over 500 generations in continuous culture of 250 days In planta. SBW25 marked at the Ee site with an integration vector (pBGS)(SBW25EeKmXylE) was equally stable.

A reliable method of transforming B. thuringiensis was developed. The segregational stability of the markers XylE and Gus, positioned on plasmid molecules under the control of Spo2 or PRLB promoters, was assessed in B. thuringiensis, Gus was unstable and therefore unsuitable, XylE and Mer were the preferred non-antibiotic markers.

Tema(s)

Data not available

Convocatoria de propuestas

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Régimen de financiación

CSC - Cost-sharing contracts

Coordinador

RIJKSUNIVERSITEIT GENT
Aportación de la UE
Sin datos
Dirección
ST PIETERNIEUWSTRAAT 25
9000 GENT
Bélgica

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Coste total
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