pMH11, A tool for gene disruption and expression analysis in Azorhizobium caulinodans.
Tools for mutagenesis and expression analyses are needed to study the role of bacterial genes. Here, we report the construction of pMH11, a small, mobilizable plasmid that replicates in Escherichia coli, but not in Azorhizobium caulinodans, a nodulating microsymbiont of Sesbania rostrata, and that contains a unique BamHI restriction site upstream of a promoterless lacZ gene. pMH11 and two derivatives with the multiple cloning site of pBluescript (KS(II)) are useful for mutagenesis by gene disruption and for expression analyses after selection for cointegration by kanamycin resistance. Weakly constitutive promoter activity from the vector allowed transcription of genes downstream of the integration site, so that no polar effects were caused by gene disruption.
D'Haeze
Wim
W
Vakgroep Moleculaire Genetica, Departement Plantengenetica, Vlaams Interuniversitair Instituut voor Biotechnologie, Universiteit Gent, K.L. Ledeganckstraat 35, B-9000 Ghent, Belgium.
Verplancke
Christa
C
Mironov
Vladimir
V
Holsters
Marcelle
M
D020599:Azorhizobium caulinodans / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D007887:Fabaceae / Q000382:microbiology; D020869:Gene Expression Profiling / Q000379:methods*; D016296:Mutagenesis; D009154:Mutation; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D013559:Symbiosis