NEW MATERIAL:Plasmid pYN32 derived from Bacillus subtilis having a restriction enzyme map expressed by the formula having about 5.4 M dalton molecular mass unit without cutting by restriction enzyme BamHI, BglI, AatI, AvaI or SalI.

USE: Used as a vector for recombinant DNA experiment.

PREPARATION: For instance, Bacillus subtilis (IFO 14132) is subjected to shaking culture in a medium and strain is collected by centrifuge, suspended in a buffer solution containing lysozyme, and allowed to stand at 37°C for 30min and centrifuged. The supernatant is mixed with isopropyl alcohol, the resultant pellet is gathered by centrifuge followed by drying, then dissolved in a buffer solution. The resultant system is mixed with a solution containing cesium chloride and rhodium bromide and subjected to ultracentrifuge. The precipitate is extracted with n-butanol and dialyzed to afford the objective plasmid pYN32.