RRC ID 33421
Author Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S.
Title Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
Journal Gene
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.
Volume 2(2)
Pages 95-113
Published 1977
PII 0378-1119(77)90000-2
PMID 344137
MeSH Ampicillin / pharmacology Conjugation, Genetic DNA, Bacterial DNA, Recombinant Escherichia coli / genetics Plasmids* Recombination, Genetic* Tetracycline / pharmacology Transformation, Bacterial
IF 2.498
Resource
Prokaryotes E. coli pBR322