Plasmids for heterologous expression in Pasteurella haemolytica.
New cloning and expression vectors that replicate both in Pasteurella haemolytica and in Escherichia coli were constructed based on a native sulfonamide (SuR) and streptomycin (SmR) resistant plasmid of P. haemolytica called pYFC1. Each shuttle vector includes an MCS and a selectable antibiotic resistance marker that is expressed in both organisms. Plasmid pNF2176 carries the P. haemolytica ROB-1 beta-lactamase gene (blaP, ApR) and pNF2214 carries the Tn903 aph3 kanamycin resistance (KmR) element. The expression vector, pNF2176, was created by placing the MCS downstream of the sulfonamide gene promoter (PsulII) on pYFC1; this was used to clone and express the promoterless Tn9 chloramphenicol resistance gene (cat, CmR) in P. haemolytica (pNF2200). A promoter-probe vector (pNF2283) was constructed from pNF2200 by deleting PsulII.
Fedorova
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Department of Microbiology and Immunology, Baylor College of Medicine, Houston, TX 77030, USA.
Highlander
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SK
D003001:Cloning, Molecular / Q000379:methods*; D004262:DNA Restriction Enzymes; D004352:Drug Resistance, Microbial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D016978:Mannheimia haemolytica / Q000235:genetics* / Q000254:growth & development; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D013307:Streptomycin; D013449:Sulfonamides
D013449:Sulfonamides; D004262:DNA Restriction Enzymes; D013307:Streptomycin