Saturation mutagenesis in Escherichia coli of a cloned Xanthomonas campestris DNA fragment with the lux transposon Tn4431 using the delivery plasmid pDS1, thermosensitive in replication.
Appl Microbiol Biotechnol
; 40(2-3): 356-60, 1993 Nov.
Article
em En
| MEDLINE
| ID: mdl-7764391
ABSTRACT
A system allowing transposon mutagenesis of cloned DNA fragments in Escherichia coli with Tn4431, which carries the promotorless luciferase (lux) operon of Vibrio fischeri, has been developed. The transposon delivery plasmid, pDS1, based on an IncF replicon, is thermosensitive in replication and mobilizable to many Gram-negative bacteria. We used pDS1 for Tn4431-saturation mutagenesis of a 10-kb DNA fragment of Xanthomonas campestris pv. campestris (X.c.c.) in E. coli and showed that the expression of the lux operon was dependent on orientation and location of the transposon. Transfer of a specific Tn4431 insertion to X.c.c. allowed the determination of the bioluminescence phenotype in planta.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA Bacteriano
/
Elementos de DNA Transponíveis
/
Mutagênese Insercional
/
Xanthomonas campestris
Idioma:
En
Revista:
Appl Microbiol Biotechnol
Ano de publicação:
1993
Tipo de documento:
Article
País de afiliação:
Alemanha