New shuttle vectors for Escherichia coil and Bacillus subtilis.

II. Plasmid pHY300PLK, a multipurpose cloning vector with a polylinker, derived from pHY460

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In vivo and in vitro recombination techniques were used to construct a new cloning vector, pHY300PLK (4.7 kb) from the shuttle vector pHY460 (7kb). The newly derived shuttle vector can replicate and express the tetracycline resistance gene (Tc^R) in both Escherichia coli and Bacillus subtilis. pHY300PLK contains the Tc^R gene, the ampicillin resistance gene (Ap^R), two replication origins for E. coli and B. subtilis and a polylinker derived from πAN7. The unique cloning sites are BalI, BamHI, BanI, BglI, BglII, BstEII, EcoRI, EcoRV, HindIII, HpaI, SalI, SmaI, PvuI and XbaI, pHY300 PLK is characterized as a copy-number mutant in E. coli. (Key words: shuttle vector; B, subtilis; tetracycline; ampicillin; copy-number mutant; incompatibility)