Pls101 contains the following restriction sites (kb markers according to the published map): PstI--0/6.4, 0.8; BclI--1.8, 2.8; HpaII--2.4; EcoRI--3.0, 4.1; HindIII--4.2, 4.9, 5.1, 5.3, 5.5, 6.2; KpnI--4.75; SacI--5.6. Cloning into the unique HpaII site inactivates tet. Removal of the smaller BclI fragment also inactivates tet. Ecori digestion removes a dispensible EcoRI fragment. Cloning into the unique KpnI and SacI sites inactivates malM. Cloning into the 4.2 kb vector fragment resulting from HindIII digestion removes malM. Mal- recombinants can be enriched by penicillin selection of tetR bacteria. Strain is noncapsulated and reportedly never reverts to encapsulated form. Strain is noncapsulated and reportedly never reverts to encapsulated form This was constructed from pLS69 (ATCC 39937) by ligating a partial 2.0 kb HindIII fragment containing malM to a 4.4 kb HindIII from a complete digest of pLS69. The latter fragment contains tet as well as the origin of replication. |
