University of California

Leland Stanford Junior University

Applied Biosystems, LLC

Dna Sciences, Inc.

The Perkin-elmer Corporation

Research, Incorporated

Alexander N Glazer

Richard A Mathies

Jingyue Ju

Scott C Benson

Lubert Stryer

Michael W Hunkapiller

Norman M Whiteley

Yuping Cai

Konan Peck

Vernon T Oi

Su-Chun Hung

Charles Cantor

Takeshi Sano

Adam T Woolley

Pankaj Singhal

John B Waterbury

Hua Xu

Kaigin Lao

Kaiqin Lao

Linda J Ong

Stanley W Watson

Aaron J Tooley

Jin Xie

Streptavidin-metallothionein chimeric proteins with biological recognition specificity in which the streptavidin moiety provides high affinity biotin binding and the metallothionein moiety provides a high affinity metal binding. The binding affinity of the streptavidin-metallothionein chimeric protein both for biotin and heavy metal ions allows specific incorporation into, conjugation with, or labelling of any biological material containing biotin with various heavy metal ions.

Streptavidin proteins

The invention provides multifunctional fusion constructs which are rapidly incorporated into a macromolecular structure such as a phycobilisome such that the fusion proteins are separated from one another and unable to self-associate. The invention provides methods and compositions for displaying a functional polypeptide domain on an oligomeric phycobiliprotein. including fusion proteins comprising a functional displayed domain and a functional phycobiliprotein domain incorporated in a functional oligomeric phycobiliprotein. The fusion proteins provide novel specific labeling reagents.

Multifunctional recombinant phycobiliprotein-based fluorescent constructs and phycobilisome display

The invention provides multifunctional fusion constructs which are rapidly incorporated into a macromolecular structure such as a phycobilisome such that the fusion proteins are separated from one another and unable to self-associate. The invention provides methods and compositions for displaying a functional polypeptide domain on an oligomeric phycobiliprotein, including fusion proteins comprising a functional displayed domain and a functional phycobiliprotein domain incorporated in a functional oligomeric phycobiliprotein. The fusion proteins provide novel specific labeling reagents.

Fluorescent labels having at least one donor and at least one acceptor fluorophore bonded to a polymeric backbone in energy transfer relationship, as well as methods for their use, are provided. Of particular interest are the subject labels wherein the polymeric backbone is a nucleic acid and the donor fluorophore is bonded to the 5' terminus of said nucleic acid. Such labels find use as primers in applications involving nucleic acid chain extension, such as sequencing, PCR and the like.

Probes labeled with energy transfer coupled dyes

Recombinant cells which express a fluorescent holo-phycobiliprotein fusion protein and methods of use are described. The cells comprises a bilin, a recombinant bilin reductase, an apo-phycobiliprotein fusion protein precursor of the fusion protein comprising a corresponding apo-phycobiliprotein domain, and a recombinant phycobiliprotein domain-bilin lyase, which components react to form the holo-phycobiliprotein fusion protein. Also described are holo-phycobiliprotein based transcription reporter cells and assays, which cells conditionally express a heterologous-to-the-cell, fluorescent, first holo-phycobiliprotein domain.

Engineering of living cells for the expression of holo-phycobiliprotein-based constructs

The present invention provides methods and kits for identifying nucleotides at a variant site in a target molecule by forming fluorescence resonance energy transfer labeled product. The location and type of fluorescent labels in the labeled product provide strong signals and relatively high spectral purity that facilitate detection. Utilizing various secondary labels and different combinations of acceptor and donor labels, certain methods permit multiple analyses to be conducted simultaneously and at high throughput. The methods can be used in a variety of applications such as analyzing point mutations and single nucleotide polymorphisms (SNPs). In addition, the methods have utility in other applications in which specific sequence information is of value, including detection of pathogens, paternity disputes, prenatal testing and forensic analysis.

Detection of nucleotide sequence variation through fluorescence resonance energy transfer label generation

Fluorescent labels having at least one donor and at least one acceptor fluorophore bonded to a polymeric backbone in energy transfer relationship, as well as methods for their use, are provided. Of particular interest are the subject labels wherein the polymeric backbone is a nucleic acid and the donor fluorophore is bonded to the 5&prime; terminus of said nucleic acid. Such labels find use as primers in applications involving nucleic acid chain extension, such as sequencing, PCR and the like.

Novel fluorescent labeling techniques and fluorescent labels are provided, employing high affinity non-covalently binding and intercalating fluorescent dyes and dsDNA. The dyes find application to provide highly sensitive labeling of nucleic acids in electrophoretic gels and as pre-prepared labels for binding to a wide variety of specific binding pair members. The DNA-dye fluorescer complex can be used for labels in diagnostic assays, detection of specific nucleic acid sequences, and the like.

Multichromophore fluorescent probes using DNA intercalation complexes

Recombinant streptavidin-metallothionein chimeric protein having biological recognition specificity

This invention relates to a microfabricated capillary electrophoresis chip for detecting multiple redox-active labels simultaneously using a matrix coding scheme and to a method of selectively labeling analytes for simultaneous electrochemical detection of multiple label-analyte conjugates after electrophoretic or chromatographic separation.

Microfabricated capillary electrophoresis chip and method for simultaneously detecting multiple redox labels

The present invention provides methods and kits for determining the identity of a nucleotide at a variant site in a target nucleic acid of interest, including, for example, point mutations and single nucleotide polymorphisms. The methods involve conducting template-dependent extension reactions in the presence of a mixture of nucleotides that include at least one labeled extendible nucleotide and at least one labeled non-extendible nucleotide that are selected to be complementary to the nucleotides that potentially occupy the variant site. The particular labeled nucleotide incorporated into the extension products is characteristic of the nucleotide at the variant site. In addition to their utility in analyzing point mutations and single nucleotide polymorphisms, the methods and kits of the invention have utility in a variety of other applications in which specific nucleotide sequence information is of value, including, for example, paternity disputes, prenatal testing, forensic analysis and detection of pathogens.

Determination of nucleotide sequence variations through limited primer extension

Compositions are provided comprising sets of fluorescent labeled oligonucleotides carrying pairs of donor and acceptor dye molecules, designed for efficient excitation of the donors at a single wavelength and emission from the acceptor in each of the pairs at different wavelengths. The different molecules having different donor-acceptor pairs can be modified to have substantially the same mobility and enhanced emission intensities under separation conditions, by varying the distance between the donor and acceptor in a given pair. Particularly, the fluorescent compositions find use as labels in analyzing double stranded and single stranded nucleic acid fragments using capillary electrophoresis.

Detection methods using probes labeled with energy transfer coupled dyes for DNA fragment analysis

Cyanine dyes are used as the donor fluorophore in energy transfer labels in which light energy is absorbed by a donor fluorophore and transferred to an acceptor fluorophore which responds to the transfer by emitting fluorescent light for detection. The cyanine dyes impart an unusually high sensitivity to the labels thereby improving their usefulness in a wide variety of biochemical procedures, particularly nucleic acid sequencing, nucleic acid fragment sizing, and related procedures.

Growing community of inventors

Orinda, CA, United States of America

Alexander N Glazer