PBR322

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Artificial plasmid

title: "PBR322" type: doc version: 1 created: 2026-02-28 author: "Wikipedia contributors" status: active scope: public tags: ["dna-mobile-genetic-elements", "molecular-biology-techniques", "plasmids"] description: "Artificial plasmid" topic_path: "science/biology" source: "https://en.wikipedia.org/wiki/PBR322" license: "CC BY-SA 4.0" wikipedia_page_id: 0 wikipedia_revision_id: 0

::summary Artificial plasmid ::

::figure[src="https://upload.wikimedia.org/wikipedia/commons/e/e8/pBR322.svg" caption="A schematic representation of the pBR322 vector with restriction sites indicated in blue."] ::

pBR322 is a plasmid and was one of the first widely used cloning vectors for E. coli. Created in 1977 in the laboratory of Herbert Boyer at the University of California, San Francisco, it was named after Francisco Bolívar Zapata, a postdoctoral researcher, and Raymond L. Rodriguez. The p stands for "plasmid," and BR for "Bolivar" and "Rodriguez."

pBR322 is 4361 base pairs in length and has two antibiotic resistance genes: blaTEM-1, which confers ampicillin resistance, and tetA, which confers tetracycline resistance. It contains the origin of replication of pMB1, and the rop gene, which encodes a restrictor of plasmid copy number. The plasmid has unique restriction sites for more than forty restriction enzymes. Eleven of these forty sites lie within the tetA gene. There are two restriction sites, HindIII and ClaI, within the promoter of the tetA gene. There are six key restriction sites inside the blaTEM-1 gene. The antibiotic resistance genes are from pSC101 for tetracycline and RSF2124 for ampicillin. The restriction sites in the antibiotic resistance genes allow for insertional inactivation.

Background

pBR322 was the most widely used early cloning vector. It has two antibiotic resistance genes as selectable markers, and over 40 unique restriction sites that made it suitable as a cloning vector. The plasmid was constructed with genetic material from three main sources: the tetracycline resistance gene of pSC101, the ampicillin resistance gene of RSF2124, and the replication elements of pMB1, a ColE1-like plasmid.

A large number of other plasmids based on pBR322 have been constructed specifically designed for a wide variety of purposes. Most expression vectors for extrachromosomal protein expression and shuttle vectors contain the pBR322 origin of replication, and fragments of pBR322 are very popular in the construction of intraspecies shuttle or binary vectors and vectors for targeted integration and excision of DNA from chromosome.

Reference samples of this plasmid are maintained by public biological resource centers, such as BCCM/GeneCorner.

DNA sequence

The sequence in pBR322 is ::data[format=table]

pBR322
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In popular culture

Books

In Michael Crichton's 1990 science fiction novel Jurassic Park “the actual structure of a small fragment of dinosaur DNA“ is shown to visitors to the Jurassic Park Research Institute. In 1992 NCBI Investigator Mark Boguski used BLAST to compare the Jurassic Park sequence with known DNA sequences, and discovered that the book sequence consisted of three sequences from pBR322, one repeated twice, separated by short random sequences. Boguski wrote up his work as a humorous paper, which was published in the journal BioTechniques.

References

References

  1. (October 1988). "A new revision of the sequence of plasmid pBR322". Gene.
  2. (November 2009). "Induction of multidrug resistance mechanism in ''Escherichia coli'' biofilms by interplay between tetracycline and ampicillin resistance genes". Antimicrobial Agents and Chemotherapy.
  3. (January 1986). "Plasmid vector pBR322 and its special-purpose derivatives — a review". Gene.
  4. (1996). "Principles of gene manipulation: an introduction to genetic engineering". Blackwell Scientific Publ.
  5. (November 1977). "Construction and characterization of new cloning vehicles I. Ampicillin-resistant derivatives of the plasmid pMB9". Gene.
  6. (November 1977). "Construction and characterization of new cloning vehicle. II. A multipurpose cloning system". Gene.
  7. (2009). ["Principles of Gene Manipulation and Genomics"](https://pharmareview.files.wordpress.com/2015/04/principle-of-gene-manipulation-and-genomics-by-sandy-b-primrose-richard-twyman.pdf }}{{Dead link). John Wiley & Sons, Ltd.
  8. (January 1985). "Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors". Gene.
  9. (April 2004). "Recombinant Gene Expression: Reviews and Protocols". Humana Press Inc..
  10. (30 September 2008). "Cloning vector pBR322, complete sequence". National Library of Medicine.
  11. (1992). "A Molecular Biologist Visits Jurassic Park". BioTechniques.
  12. (9 December 2018). "Post-Publication Review of Jurassic Park by Michael Crichton". [[Dan Graur]].

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dna-mobile-genetic-elementsmolecular-biology-techniquesplasmids