Daniel B Simpson

US Patent:

52083322, May 4, 1993

Filed:

Dec 20, 1991

Appl. No.:

7/811217

Inventors:

Babetta L. Marrone - Los Alamos NM
Daniel J. Simpson - Los Alamos NM
Clifford J. Unkefer - Los Alamos NM
Thomas W. Whaley - Santa Fe NM

Assignee:

The United States of America as represented by the United States
Department of Energy - Washington DC

International Classification:

C07D26538

US Classification:

544102

Abstract:

An optical probe enables the study of enzyme activity by absorbance spectroscopy or by sensitive fluorescence methods. In particular, the probe provides the ability to monitor the activity of cytochrome P-450. sub. scc enzyme, the rate limiting enzyme for steroid biosynthesis. Located on the inner mitochondrial membrane, P-450. sub. scc catalyzes the conversion of cholesterol to pregnenolone and isocapraldehyde by sequential oxidations of the cholesterol side chain. The fluorogenic probe includes a cholesterol-like steroid linked to a chromophore through a linking group. The chromophore is selected to have little optical response when linked to the steroid substrate and an enhanced optical response when cleaved from the substrate and linking group. Thus, a fluorescent anion that can be optically detected is generated by the side-chain cleavage reaction during steroidogenesis.

US Patent:

54057477, Apr 11, 1995

Filed:

Mar 7, 1994

Appl. No.:

8/208506

Inventors:

James H. Jett - Los Alamos NM
Richard A. Keller - Los Alamos NM
John C. Martin - Los Alamos NM
Richard G. Posner - Los Alamos NM
Babetta L. Marrone - Los Alamos NM
Mark L. Hammond - Los Alamos NM
Daniel J. Simpson - Los Alamos NM

Assignee:

The Regents of the University of California Office of Technology Transfer - Alameda CA

International Classification:

C12Q 168
C12P 1934
G01N 3348

US Classification:

435 6

Abstract:

Method for rapid-base sequencing in DNA and RNA with two-base labeling and employing fluorescent detection of single molecules at two wavelengths. Bases modified to accept fluorescent labels are used to replicate a single DNA or RNA strand to be sequenced. The bases are then sequentially cleaved from the replicated strand, excited with a chosen spectrum of electromagnetic radiation, and the fluorescence from individual, tagged bases detected in the order of cleavage from the strand.

US Patent:

51107252, May 5, 1992

Filed:

Apr 5, 1991

Appl. No.:

7/680978

Inventors:

Babetta L. Marrone - Los Alamos NM
Daniel J. Simpson - Los Alamos NM
Clifford J. Unkefer - Los Alamos NM
Thomas W. Whaley - Santa Fe NM

Assignee:

The United States of America as represented by the United States
Department of Energy - Washington DC

International Classification:

C12Q 160
C12N 902

US Classification:

435 11

Abstract:

An optical probe enables the study of enzyme activity by absorbance spectroscopy or by sensitive fluorescence methods. In particular, the probe provides the ability to monitor the activity of cytochrome P-450. sub. scc enzyme, the rate limiting enzyme for steroid biosynthesis. Located on the inner mitochondrial membrane, P-450. sub. scc catalyzes the conversion of cholesterol to pregnenolone and isocapraldehyde by sequential oxidations of the cholesterol side chain. The fluorogenic probe includes a cholesterol-like steroid linked to a chromophore through a linking group. The chromophore is selected to have little optical response when linked to the steroid substrate and an enhanced optical response when cleaved from the substrate and linking group. Thus, a fluorescent anion that can be optically detected is generated by the side-chain cleavage reaction during steroidogenesis.