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METHODS Media and materials. All the media used in this work have been described by Imanaka et a/. (1 98 I a, b, 1982). The companies and laboratories, from which the antibiotics, restriction enzymes and all other reagents used here were purchased were the same as those used in previous works (Imanaka et a/., 1981a, b, 1982). Bacterial strain and plasmids. Bacillus stearothermophilus CU21 (streptomycin resistant ; Imanaka ef at., 1982) was used as the host strain. pTB90 (encoding resistance to both kanamycin and tetracycline, Km' Tc'; Imanaka rt a/., 1982) was used as the vector plasmid. Penicillinase genes from both the constitutive and wild-type strains of B. lichcnijormis 9945A have been cloned in Escherzchia coli with pMB9 (Tc') as vector plasmid. These recombinant plasmids are designated, pTTE 1 1 (Tc' penP+ pmllO) and pTTE2 1 (Tc' penP+ peril+), respectively (Imanaka et ul., 1981 h). penP and penlare the structural gene and the repressor gene for penicillinase, respectively, and are coded on the same EcoRI fragment (2-8 MDal, Imanaka et a/., 1981 b). The restriction endonuclease cleavage maps of these plasmids are shown in the upper row of Fig. I.

Molecular Cloning and Expression of Penicillinase Genes from Bacillus licheniformis in the Thermophile Bacillus stearothermophilus