[Use of a plasmid with integrative function of phage phiC31 for transfer of cloned genes into Streptomyces strains]

Opportunities for application of integrative vectors carrying the attP site and the int gene of the temperate actinophage phi C31 for cloning genes in Streptomyces strains were demonstrated. The behavior of the integrative vectors pZAT22 and pTO1 in the model strain S. lividans TK64 and in the bialaphos-producing strain S. hygroscopicus, respectively, was characterized. Restriction maps of the S. lividans and S. hygroscopicus chromosomal regions containing attB sites were constructed. The bar gene of resistance to bialaphos was incorporated into the chromosome of the model strain S. lividans via the integrative pZAT22 vector, the level of expression of this gene within the chromosome of a heterologous host was determined. The possibility of amplification of the bar gene in the chromosome of the strain S. lividans was shown. The conjugative integrative vector pTO1, which carried the cloned bar gene, was introduced into the bialaphos-producing strain by intergeneric matings of Escherichia coli and S. hygroscopicus. The effect of an additional copy of the gene in the chromosome of S. hygroscopicus on strain resistance and productivity was studied.