Novel 2 to 4 Base Pair Specific Enzymes for PCR Fingerprinting

Award Information
Agency: Department of Agriculture
Branch: N/A
Contract: N/A
Agency Tracking Number: 34397
Amount: $150,000.00
Phase: Phase II
Program: SBIR
Awards Year: 1997
Solicitation Year: N/A
Solicitation Topic Code: N/A
Solicitation Number: N/A
Small Business Information
Megabase Research Products
2820 North 48th Street, Lincoln, NE, 68504
DUNS: N/A
HUBZone Owned: N
Woman Owned: N
Socially and Economically Disadvantaged: N
Principal Investigator
 Dr. Michael Nelson
 Principal Investigator
 () -
Business Contact
Phone: () -
Research Institution
N/A
Abstract
A large number of RFLP variants in plant and animal diseases can be detected using the polymerase chain reaction (PCR) followed by restriction endonuclease cleavage. However, the resolution of PCR fingerprinting techniques is limited by rather long (4 to 6 base pair) DNA sequence specificities of most bacterial restriction enzymes. Bacterail restriction sites occur, on average, once every 4^4 to 4^6 b.p.; and most such sites are not polymophic. In contrast, Chlorella virus (Cvi) endonucleases have short (2 to 4 base pair) sequence specificities and therefore ideal for find structure gene mapping applications such as PCR-coupled mapping. Cvi enzymes cut once every 4^2 to 4^4 b.p. provide a simple, inexpensive means of rapidly fingerprinting genetic variants of economically important plant and animal diseases. Phase I studies are proposed to purify five different Cvi endonucleases which have short (2 to 3 base pair) sequence specificities: CviJI (RG CY), CviNYSI (CC), Cvi109I (YC GR), CviNY2AI (R AG), and CviGI (CGR). These frequently cutting Cvi endonucleases will be tested in PCR fingerprinting of an agriculturally important viral plant pathogen, Barley Yellow Dwarf Virus (BYDV)Cvi endonucleases represent a major new class of fine structure gene mapping tool.

* information listed above is at the time of submission.

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