Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
In vitro recombination techniques were used to construct a new cloning vehicle, pBR322. This plasmid, derived from pBR313, is a relaxed replicating plasmid, does not produce and is sensitive to colicin E1, and carries resistance genes to the antibiotics ampicillin (Ap) and tetracycline (Tc). The antibiotic-resistant genes on pBR322 are not transposable. The vector pBR322 was constructed in order to have a plasmid with a single PstI site, located in the ampicillin-resistant gene (Apr), in addition to four unique restriction sites, EcoRI, HindIII, BamHI and SalI. Survival of Escherichia coli strain X1776 containing pBR313 and pBR322 as a function of thymine and diaminopimelic acid (DAP) starvation and sensitivity to bile salts was found to be equivalent to the non-plasmid containing strain. Conjugal transfer of these plasmids in bi- and triparental matings were significantly reduced or undetectable relative to the plasmid ColE1.
D000667:Ampicillin / Q000494:pharmacology; D003227:Conjugation, Genetic; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics; D010957:Plasmids; D011995:Recombination, Genetic; D013752:Tetracycline / Q000494:pharmacology; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D004274:DNA, Recombinant; D000667:Ampicillin; D013752:Tetracycline