FIG. 1.

Flowchart of construction of L protein expression plasmid. The L gene was amplified in three fragments by PCR. The primers are indicated above each fragment. Restriction enzyme recognition sites used for cloning are indicated. Sites used for assembly of pUC-L and subcloning of the L gene into pCITE-L are marked by arrows. At the bottom, details of the subcloning of the L gene into the NcoI site of pCITE-L are shown. Note that cloning was possible, although the overhang at the 5′ end of L gene was not fully compatible with an NcoI overhang.