Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.

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.

• Authors: F Bolivar, R L Rodriguez, P J Greene, M C Betlach, H L Heyneker, H W Boyer, J H Crosa, S Falkow
• Journal: Gene (Gene) Vol. 2 Issue 2 Pg. 95-113 ( 1977) ISSN: 0378-1119 [Print] Netherlands
• PMID: 344137 (Publication Type: Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA, Bacterial
• DNA, Recombinant
• Ampicillin
• Tetracycline

Topics

• Ampicillin (pharmacology)
• Conjugation, Genetic
• DNA, Bacterial
• DNA, Recombinant
• Escherichia coli (genetics)
• Plasmids
• Recombination, Genetic
• Tetracycline (pharmacology)
• Transformation, Bacterial