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ANNETTE D E I CHMANN AND KLAUS DEICHMANN
cos site
cos site
cos site
cos site
restriction
digest
cos site
~tion
cos site
genomic DNA
about 40.000 bp
recombinant plasmids forming concatemeres
+ helper phage
bacteriophages
~
transfection
hostcett
cos site
Fig. 4. Cosmids use the transfection mechanisms ofbacteriophage to enter cells. Once they
are intracellular they are amplified like plasmids (mcr = multiple cloning region).
Subprotocol 1
Transformation of Bacteria with Recombinant Plasmid Using the Ca++Precipitation Method (Mandel 1970)
The bacterial strain tobe transformed depends on the selected vector. For
pBR322 and pBR322-derived vectors, E. coli HBlOl is a commonly used
bacterial strain. The following transformation procedure refers to this
strain. If you wish to use a different strain, the principles of the method