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Engineering New Restriction Endonucleases

Award Information
Agency: Department of Health and Human Services
Branch: N/A
Contract: 1 R43 GM52258-01,
Agency Tracking Number: 29449
Amount: $680,165.00
Phase: Phase II
Program: SBIR
Solicitation Topic Code: N/A
Solicitation Number: N/A
Timeline
Solicitation Year: N/A
Award Year: 1996
Award Start Date (Proposal Award Date): N/A
Award End Date (Contract End Date): N/A
Small Business Information
32 Tozer Road
Beverly, MA 01915
United States
DUNS: N/A
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 Ira Schildkraut
 () -
Business Contact
Phone: () -
Research Institution
N/A
Abstract

Restriction endonucleases are of great importance to molecular DNA cloning, DNA diagnostics and genengineering. There is an ever increasing demand for a larger repertoire of recognition specificitiesendonuclease field is at point where the engineering of cleavage specificities is approachable. BamHfor engineering new specificities. It has been cloned, overexpressed and crystallized. BamHI bindingdeficient (cat-) variants have been isolated by using an in vivo transcriptional interference selectthat only cats variants can lead to spectinomycin resistance (spR). By inserting the BamHI recognitiinto an antisense promoter, transcription can be regulated. Binding of the cats BamHI to this operatthereby relieving the transcriptional interference resulting in spR. The 1.95 Angstroms resolution tstructure of BamHI has led to a determination of four amino acid residues within a seven amino acidrecognition. Cassette mutagenesis which varies all four DNA recognizing residues of a cats BamHI varrelatively few mutants (-160,000). Among these mutants, those which bind to anti-sense promoter operthan GGATCC can easily be selected. After selection of altered binding variants, cleavage will be recar mutation.

* Information listed above is at the time of submission. *

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