Construction of a vector plasmid family and its use for molecular cloning in Streptococcus lactis.

ABSTRACT

Cloning vector plasmids have been constructed on the basis of the broad host range plasmid pAM beta 1 and used for the cloning of a nisin resistance determinant in Streptococcus lactis. They incorporate several desirable features for gene cloning in S. lactis and other transformable Gram-positive bacteria. They carry an easily selectable erythromycin resistance marker, are present at low (6-9) or high (45-85) copy number in S. lactis and possess a convenient polyrestriction site sequence. A significant advantage of these plasmids is their capability to carry and stably maintain very large cloned DNA fragments (up to 30 kilobases).