1887

Abstract

Transposon Tn was used to generate a fructokinase mutation in biovar BAL. The section of the genome containing Tn was cloned into the RI site of the vector pHC79 and isolated by direct selection on medium containing kanamycin and tetracycline. Total RI digestion was used to obtain a single fragment containing Tn and flanking DNA sequences. The flanking DNA was used as a probe to isolate an intact fructokinase gene from a pLAFRl cosmid clone bank of the parental strain. A cosmid showing homology to the probe was tri-parentally conjugated into the fructokinase-negative strain, complementing the mutation. The complemented mutant exhibited the wild-type phenotype, with an increase in fructokinase production presumably due to multiple copies of the gene.

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1989-08-01
2021-10-20
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