Gene Mutation Technique For Producing Strain Of The Single Component Of Apramycin

Streptomyces tenebrarius H6 produces three kinds of antibiotics: ampramycin, carbamoyltobramycin and some carbamoylkcanamycinB. To study the biosynthetic pathway of these antibiotics and obtain the S.tenebrarius which produce ampramycin only, the tacA has been disrupted. First, the 1063bp and 523bp inside fragments of tacA gene was obtained by PCR, the gene disruption vector (pSPU292) was constructed and transferred into S.tenebrarius H6 by conjugation. Recombinant S.tenebrarius H6(pSPU292)was obtained. With producing apramycin, the disruptant no longer produces carbamoyltobramycin ,the result indicates that the tacA is not related to apramycin biosynthesis but closely related to carbamoyltobramycin biosynthesis.