| WARF: P98090US |  |
| Conditionally Amplifiable BAC Vectors |
| INVENTORS | • | Waclaw Szybalski, Jadwiga Wild, Zdenka Hradecna |
OVERVIEW
Because they retain large inserts over many generations, bacterial artificial chromosomes (BACs) are currently the vectors of choice for maintaining clones of large genomic DNA fragments. However, because each BAC clone exists as only a single copy within host cells,
large numbers of cells must be grown to generate enough material for analyses such as
sequencing. This invention provides a simple but highly controllable system for amplifying DNA inserts from BAC vectors. This improved BAC
vector contains a pair of excision-mediating sites (EMS) that flank a cloning site for foreign DNA, and an origin of replication (ori) site. After an appropriate host cell is transformed with an insert-containing BAC, the cell can be induced to produce a protein that excises the DNA region between the EMS from the BAC. The excised fragment is then circularized, creating a plasmid that contains both the insert and the ori. Upon exposure to a second induction event, the host cell initiates replication at the ori, so that the insert-containing plasmid is amplified and accumulates in high number (typically 10 to 500 copies). Methods for constructing these improved BAC vectors are provided.
KEY BENEFITS
- BAC has the capacity not only to effectively maintain large-size, cloned DNA fragments, but also to directly yield multiple copies of these inserts without the need for additional cloning steps
- Excision and amplification of DNA inserts are tightly controlled by the presence of appropriate signals
- Large-size, cloned DNA fragments may be amplified 10-500 times without the need for PCR methods
- Methods are applicable to any BAC vector that is capable of independent replication in host cells
- Especially suited for genome sequencing efforts because of the large insert capacity of the BAC and the closely controllable nature of the amplification system
ADDITIONAL INFORMATION
Related Technologies
Intellectual Property Status
Tech Fields
Research Tools - DNA & RNA tools
Research Tools - Genomics & proteomics
CONTACT INFORMATION
For current licensing status, please contact our team at
licensing@warf.org
or phone 608.262.4924. (Clicking this link will open a contact form in a popup window. If you have problems viewing the form, try disabling your popup blocker software.)
Since its founding in 1925 as the patenting and licensing organization for the University of Wisconsin-Madison, WARF has been working with business and industry to transform university research into products that benefit society. WARF intellectual property managers and licensing staff members are leaders in the field of university-based technology transfer. They are familiar with the intricacies of patenting, have worked with researchers in relevant disciplines, understand industries and markets, and have negotiated innovative licensing strategies to meet the individual needs of business clients.
