Construction of a single-transposon-insertion mutant in Rhizobium sp. strain TAL1145 from a double-insertion mutant.
A method for developing a single-transposon-insertion mutant from a double-insertion mutant in Rhizobium is described. An exopolysaccharide (EPS)-defective mutant containing two Tn5-lacZ insertions was complemented with cloned wild-type DNA for EPS synthesis. One of the Tn5-lacZ insertions from the mutant was transferred to the complementing plasmid by homologous recombination. The plasmid containing the Tn5-lacZ insertion in the gene involved in EPS synthesis was transferred into the wild-type strain and the Tn5-lacZ was homogenized to obtain an EPS-defective mutant with a single Tn5-lacZ insertion.
Parveen
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N
Department of Plant Molecular Physiology, University of Hawaii, Honolulu 96822.
Borthakur
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D
D015139:Blotting, Southern; D004251:DNA Transposable Elements / Q000235:genetics; D016254:Mutagenesis, Insertional / Q000379:methods*; D010957:Plasmids; D017354:Point Mutation / Q000235:genetics*; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis / Q000235:genetics*; D012231:Rhizobium / Q000235:genetics*; D014162:Transfection
D004251:DNA Transposable Elements; D011135:Polysaccharides, Bacterial