Novel 2 to 4 Base Pair Specific Enzymes for PCR Fingerprinting

Award Information
Agency:
Department of Agriculture
Branch
n/a
Amount:
$150,000.00
Award Year:
1997
Program:
SBIR
Phase:
Phase II
Contract:
n/a
Award Id:
34397
Agency Tracking Number:
34397
Solicitation Year:
n/a
Solicitation Topic Code:
n/a
Solicitation Number:
n/a
Small Business Information
2820 North 48th Street, Lincoln, NE, 68504
Hubzone Owned:
N
Minority Owned:
N
Woman Owned:
N
Duns:
n/a
Principal Investigator:
Dr. Michael Nelson
Principal Investigator
() -
Business Contact:
() -
Research Institute:
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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