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.

Keywords

MembraneBovine serum albuminBiologyEscherichia coliMinimum inhibitory concentrationAntimicrobialCloning (programming)ChromatographyBiochemistryChemistryMicrobiologyGene

MeSH Terms

AmpicillinConjugationGeneticDNABacterialDNARecombinantEscherichia coliPlasmidsRecombinationGeneticTetracyclineTransformationBacterial

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Publication Info

Year
1977
Type
article
Volume
2
Issue
2
Pages
95-113
Citations
3854
Access
Closed

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Cite This

Francisco Bolívar, Raymond L. Rodriguez, Patricia J. Greene et al. (1977). Construction and characterization of new cloning vehicle. II. A multipurpose cloning system. Gene , 2 (2) , 95-113. https://doi.org/10.1016/0378-1119(77)90000-2

Identifiers

DOI
10.1016/0378-1119(77)90000-2
PMID
344137

Data Quality

Data completeness: 81%