PMID:344137

Citation	Bolivar, F, Rodriguez, RL, Greene, PJ, Betlach, MC, Heyneker, HL, Boyer, HW, Crosa, JH and Falkow, S (1977) Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95-113
Abstract	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.
Links	PubMed
Keywords	Ampicillin/pharmacology; Conjugation, Genetic; DNA, Bacterial; DNA, Recombinant; Escherichia coli/genetics; Plasmids; Recombination, Genetic; Tetracycline/pharmacology; Transformation, Bacterial
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