Life Technologies Corporation

Invitrogen Corporation

Biogrammatics, Inc.

Sloan-kettering Institute for Cancer Research

South Carolina Research Institute

Knut R Madden

Jonathan D Chesnut

John J Carrino

James Fan

Martin Anthony Gleeson

Louis Leong

James L Hartley

Michael A Brasch

David Cheo

Gary F Temple

Robert Bennett

James M Cregg

Thomas G Chappell

Ilya I Tolstorukov

Devon R Byrd

John A Heyman

Stewart Shuman

Devon R N Byrd

Jonathan Chestnut

Michael Felder

Adam Harris

Gabor Szalai

Martin A G Gleeson

Devon Rn Byrd

Adam N Harris

Gray F Temple

Miroslav Dudas

Bennett Robert

John Heyman

Steward Shuman

Byung-In Lee

Robert P Bennett

Karl H Hecker

In accordance with the invention, isolated nucleic acids, expression methods, host cells, expression vectors, and DNA constructs for producing proteins, and proteins produced using the expression methods are described. More particularly, nucleic acids isolated fromwherein the nucleic acids have promoter activity are described. The invention also relates to expression methods, host cells, expression vectors, and DNA constructs, for using thepromoters to produce proteins and polypeptides, and to the proteins and polypeptides produced using the expression methods.

Yeast promotors for protein expression

Isolated nucleic acids, expression methods, host cells, expression vectors, and DNA constructs for producing proteins, and proteins produced using the expression methods are disclosed. More specifically, nucleic acids isolated frompastons having promoter activity and expression methods, host cells, expression vectors, and DNA constructs of using thepastons promoters to produce different proteins and polypeptides are disclosed.

Yeast promoters for protein expression

The present invention provides compositions and methods for recombinational cloning. The compositions include vectors having multiple recombination sites and/or multiple topoisomerase recognition sites. The methods permit the simultaneous cloning of two or more different nucleic acid molecules. In some embodiments the molecules are fused together while in other embodiments the molecules are inserted into distinct sites in a vector. The invention also generally provides for linking or joining through recombination a number of molecules and/or compounds (e.g., chemical compounds, drugs, proteins or peptides, lipids, nucleic acids, carbohydrates, etc.) which may be the same or different. The invention also provides host cells comprising nucleic acid molecules of the invention or prepared according to the methods of the invention, and also provides kits comprising the compositions, host cells and nucleic acid molecules of the invention, which may be used to synthesize nucleic acid molecules according to the methods of the invention.

Methods and compositions for synthesis of nucleic acid molecules using multiple recognition sites

Methods and compositions for synthesis of nucleic acid molecules using multiplerecognition sites

The present invention provides compositions, methods, and kits for covalently linking nucleic acid molecules. The methods include a strand invasion step. For example, a method of covalently linking double stranded (ds) nucleic acid molecules can include contacting a first ds nucleic acid molecule, which has a topoisomerase linked to a 3' terminus of one end and has a single stranded 5&#x2032; overhang at the same end, with a second ds nucleic acid molecule having a blunt end, such that the 5&#x2032; overhang can hybridize to a complementary sequence of the blunt end of the second nucleic acid molecule, and the topoisomerase can covalently link the ds nucleic acid molecules. The methods are simpler and more efficient than previous methods for covalently linking nucleic acid sequences, and the compositions and kits facilitate practicing the methods, including methods of directionally linking two or more ds nucleic acid molecules.

Methods and reagents for molecular cloning

The present invention is in the fields of biotechnology and molecular biology. More particularly, the present invention relates to cloning or subcloning one or more nucleic acid molecules comprising one or more type IIs restriction enzyme recognition sites. The present invention also embodies cloning such nucleic acid molecules using recombinational cloning methods such as those employing recombination sites and recombination proteins. The present invention also relates to nucleic acid molecules (including RNA and iRNA), as well as proteins, expressed from host cells produced using the methods of the present invention.

Methods and compositions for seamless cloning of nucleic acid molecules

A method of generating a double stranded (ds) recombinant nucleic acid molecule covalently linked in both strands by contacting two or more ds nucleotide sequences with a topoisomerase under conditions such that both termini of at least one end of a first ds nucleotide sequence are covalently linked by the topoisomerase to both termini of at least one end of a second ds nucleotide sequence is provided. Also provided is a method for generating a ds recombinant nucleic acid molecule covalently linked in one strand, by contacting two or more ds nucleotide sequences with a type IA topoisomerase under conditions such that one strand, but not both strands, of one or both ends of a first ds nucleotide sequence are covalently linked by the topoisomerase. Compositions for performing such methods, and compositions generated from such methods also are provided, as are kits containing components useful for conveniently practicing the methods.

Methods and compositions for generating recombinant nucleic acid molecules

The present invention provides compositions, methods, and kits for covalently linking nucleic acid molecules. The methods include a strand invasion step, and the compositions and kits are useful for performing such methods. For example, a method of covalently linking double stranded (ds) nucleic acid molecules can include contacting a first ds nucleic acid molecule, which has a topoisomerase linked to a 3' terminus of one end and has a single stranded 5&#x2032; overhang at the same end, with a second ds nucleic acid molecule having a blunt end, such that the 5&#x2032; overhang can hybridize to a complementary sequence of the blunt end of the second nucleic acid molecule, and the topoisomerase can covalently link the ds nucleic acid molecules. The methods are simpler and more efficient than previous methods for covalently linking nucleic acid sequences, and the compositions and kits facilitate practicing the methods, including methods of directionally linking two or more ds nucleic acid molecules.

A Vaccinia topoisomerase-adapted linker, which contains an oligonucleotide primer binding site of known sequence, useful for specifically joining the linker to the end of a polynucleotide of unknown sequence and allowing subsequent PCR amplification of the polynucleotide or DNA is provided. Kits containing the invention Vaccinia topoisomerase-adapted linker and one or more linker-specific oligonucleotides for annealing to the linker in PCR are also provided. In addition, the invention provides methods for using the invention linkers in linker-mediated PCR amplification procedures for isolation and optional sequencing of isolated PCR amplification products.

Topoisomerase linker-mediated amplification methods

Compositions and methods for rapidly generating recombinant nucleic acid molecules

Growing community of inventors

Carlsbad, CA, United States of America

Knut R Madden