Electrotransformation of Lactobacillus plantarum using linearized plasmid DNA.
Evidence is presented that linearized plasmid DNA is capable of electrotransforming Lactobacillus plantarum at a frequency 500-fold lower than with the covalently closed circular molecule. When the linearized plasmid was religated prior to transformation the transformation efficiency was < 10-fold higher, suggesting that open circular plasmid was only slightly more efficient in the transformation of Lact. plantarum than linear DNA. This observation has implications for direct cloning into this species since the high background transformation frequency produced by the linear DNA could potentially obscure the recovery of clones. Nevertheless, using positive selection for enhanced chloramphenicol resistance, cloned fragments of Lact. helveticus DNA were obtained using the shuttle vector pGKV110.
Thompson
J K
JK
Food Science Division (Food Microbiology), Department of Agriculture for Northern Ireland, Belfast, UK. kthompson@alpha1.dani.gov.uk
McConville
K J
KJ
McReynolds
C
C
Collins
M A
MA
D007778:Lactobacillus / Q000235:genetics*; D010957:Plasmids; D014169:Transformation, Bacterial