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Wall-deficient forms of fast-growing mycobacteria were produced in growth medium containing vancomycin and glycine, and spheroplasts were prepared by lysozyme treatment of wall-deficient cells. Spheroplasts gave rise to recombinants with high frequency (2-6%) when they were fused using polyethylene glycol 6000. The results demonstrated that in vivo genetic recombination could be used to produce genetically modified Mycobacterium strains with applications in transformation of steroids. Useful intermediates of steroid drug synthesis and new degradation products were obtained from sterols by selected recombinant strains.
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