Improving transformation rate for Bacillus licheniformis host - by incubating foreign DNA with host cell modifying enzyme before transfer

摘要

The number of transformants produced when transferring foreign DNA into a receptor microorganism is increased by (1) releasing restriction and modification enzymes from a receptor (Bacillus licheniformis) cell; (2) inhibiting the restriction enzymes: (3) the modification system is allowed to act in vitro on the foreign DNA in presence of a c-factor; then (4) transferring the modified DNA by standard methods into the same receptor organism. Also new are (A) cell lysates with methylase activity obtd. from B. licheniformis DSM 641,3406 and/or 3407 (or their mutants or variants) by growing the cells mechanically or enzymatically disintergrating them, sepn. of large components or high mol. wt. materials by filtration and/or centrifugation, and (B) isolated DNA contg., at least once, the recognition site 5'-GCNGC-3', which is methylated in vitro by the cell lysate. USE/ADVANTAGE - Provides improved succes rate in B. licheniformis transformants.