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Novel 2 to 4 Base Pair Specific Enzymes for PCR Fingerprinting

Award Information

Department of Agriculture
Award ID:
Program Year/Program:
1997 / SBIR
Agency Tracking Number:
Solicitation Year:
Solicitation Topic Code:
Solicitation Number:
Small Business Information
Megabase Research Products
4711 Huntington Ave., Suite 2W Lincoln, NE 68504
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Woman-Owned: Yes
Minority-Owned: No
HUBZone-Owned: No
Phase 2
Fiscal Year: 1997
Title: Novel 2 to 4 Base Pair Specific Enzymes for PCR Fingerprinting
Agency: USDA
Contract: N/A
Award Amount: $150,000.00


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.

Principal Investigator:

Dr. Michael Nelson
Principal Investigator

Business Contact:

Small Business Information at Submission:

Megabase Research Products
2820 North 48th Street Lincoln, NE 68504

Number of Employees: N/A
Woman-Owned: No
Minority-Owned: No
HUBZone-Owned: No