Site-specific Chromosomal Integration Mediated By PhiC31 Integrase

Clipboard, Search History, and several other advanced features are temporarily unavailable. Skip to main page content

The .gov means it’s official. Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

The site is secure. The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation Search: Search Advanced Clipboard Save Email Send to

• Clipboard
• My Bibliography
• Collections
• Citation manager

Display options Display options Format Abstract PubMed PMID

Save citation to file

Format: Summary (text) PubMed PMID Abstract (text) CSV Create file Cancel

Email citation

Email address has not been verified. Go to My NCBI account settings to confirm your email and then refresh this page. To: Subject: Body: Format: Summary Summary (text) Abstract Abstract (text) MeSH and other data Send email Cancel

Add to Collections

• Create a new collection
• Add to an existing collection

Name your collection: Name must be less than 100 characters Choose a collection: Unable to load your collection due to an error Please try again Add Cancel

Add to My Bibliography

• My Bibliography

Unable to load your delegates due to an error Please try again Add Cancel

Your saved search

Name of saved search: Search terms: Test search terms Would you like email updates of new search results? Saved Search Alert Radio Buttons

• Yes
• No

Email: (change) Frequency: Monthly Weekly Daily Which day? The first Sunday The first Monday The first Tuesday The first Wednesday The first Thursday The first Friday The first Saturday The first day The first weekday Which day? Sunday Monday Tuesday Wednesday Thursday Friday Saturday Report format: Summary Summary (text) Abstract Abstract (text) PubMed Send at most: 1 item 5 items 10 items 20 items 50 items 100 items 200 items Send even when there aren't any new results Optional text in email: Save Cancel

Create a file for external citation management software

Create file Cancel

Your RSS Feed

Name of RSS Feed: Number of items displayed: 5 10 15 20 50 100 Create RSS Cancel RSS Link Copy

Full text links

Springer Full text links

Actions

CiteCollectionsAdd to Collections

• Create a new collection
• Add to an existing collection

Name your collection: Name must be less than 100 characters Choose a collection: Unable to load your collection due to an errorPlease try again Add Cancel PermalinkPermalinkCopyDisplay options Display options Format AbstractPubMedPMID

Page navigation

• Title & authors
• Abstract
• Publication types
• MeSH terms
• Substances
• Grants and funding
• LinkOut - more resources

Title & authors Abstract Publication types MeSH terms Substances Grants and funding LinkOut - more resources Full text links CiteDisplay options Display options Format AbstractPubMedPMID

Abstract

phiC31 integrase is a site-specific recombinase from a bacteriophage that has become a useful tool in mammalian cells. The enzyme normally performs precise, unidirectional recombination between two attachment or att sites called attB and attP. We have shown that an attP site preintegrated into a mammalian chromosome can serve as a target for integration of an introduced plasmid carrying an attB site. Recombination leads to precise integration of the plasmid into the chromosome at the attP site. This reaction is useful for placing introduced genes into the same chromosomal environment, in order to minimize position effects associated with random integration. Because phiC31 integrase can also mediate integration at endogenous sequences that resemble attP, called pseudo attP sites, a selection system is used that yields integration only at the desired preintegrated attP site. This chapter provides a protocol that features a simple antibiotic selection to isolate cell lines in which the introduced plasmid has integrated at the desired attP site. A polymerase chain reaction assay is also presented to verify correct chromosomal placement of the introduced plasmid. This integration system based on phiC31 integrase supplies a simple method to obtain repeated integration at the same chromosomal site in mammalian cells.

PubMed Disclaimer

Publication types

• Research Support, N.I.H., Extramural Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search

MeSH terms

• Attachment Sites, Microbiological Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Bacteriophages / enzymology Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Bacteriophages / genetics Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Cell Line Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Genetic Techniques Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Humans Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Integrases / genetics* Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Integrases / metabolism Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Plasmids / genetics Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search
• Polymerase Chain Reaction Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search

Substances

• Integrases Actions
  • Search in PubMed
  • Search in MeSH
  • Add to Search

Grants and funding

• EY16702/EY/NEI NIH HHS/United States
• HL68112/HL/NHLBI NIH HHS/United States

LinkOut - more resources

• Full Text Sources

  • Springer

• Other Literature Sources

  • The Lens - Patent Citations Database

Full text links [x] Springer [x] Cite Copy Download .nbib .nbib Format: AMA APA MLA NLM Send To

• Clipboard
• Email
• Save
• My Bibliography
• Collections
• Citation Manager

[x]

NCBI Literature Resources

MeSH PMC Bookshelf Disclaimer

The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Unauthorized use of these marks is strictly prohibited.