Vector Database

A BamHI digest removes a 930 bp fragment containing the lacZalpha coding frame. Thus use of BamHI as a cloning site and selection of clear plaques on Xgal media may not indicate the presence of a recombinant insert. lambdaDL10 (ATCC 37489) and lambdaDL11 (ATCC 37490) have opposite orientations of the restriction sites within lacZ. A 435 bp HaeII fragment of pUC9, encoding beta-galactosidase, was inserted into the HindIII site of lambdaD69 to produce lambdaDL9. The multiple cloning sites of M13mp10 were crossed into lambdaDL9 to produce lambdaDL10. E.coli JM109 is not a good host for this vector. (personal communication) If HindIII, SacI, XbaI, XhoI, or BamHI are used as cloning sites, the vector can accept fragments of 0 - 12 kb. Inserts in HindIII, SacI, XbaI, BamHI, and SalI inactivate lacZ. If SalI is used as a cloning site, the vector can accept fragments of 9 - 20 kb. Replacement of the SalI fragment with a DNA insert results in a lambdacIII- phage that forms virtually clear plaques. Medium is 1227 LB plus ampicillin. Hosts: bacteria-free lysate, E.coli C600, E.coli JM101. (Information source: VectorDB ( http://seq.yeastgenome.org/vectordb ).)