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1. WO1996002660 - POSITIONAL CONTROL OF SELENIUM INSERTION IN POLYPEPTIDES FOR X-RAY CRYSTALLOGRAPHY

Publication Number WO/1996/002660
Publication Date 01.02.1996
International Application No. PCT/US1995/009121
International Filing Date 19.07.1995
Chapter 2 Demand Filed 20.02.1996
IPC
C12N 9/08 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
02Oxidoreductases (1.), e.g. luciferase
08acting on hydrogen peroxide as acceptor (1.11)
C12N 15/10 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
C12P 21/00 2006.01
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
21Preparation of peptides or proteins
G01N 23/20 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
23Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/-G01N17/178
20by using diffraction of the radiation by the materials, e.g. for investigating crystal structure; by using scattering of the radiation by the materials, e.g. for investigating non-crystalline materials; by using reflection of the radiation by the materials
CPC
A01K 2217/05
AHUMAN NECESSITIES
01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
KANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
2217Genetically modified animals
05Animals comprising random inserted nucleic acids (transgenic)
A01K 67/0275
AHUMAN NECESSITIES
01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
KANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
67Rearing or breeding animals, not otherwise provided for; New breeds of animals
027New breeds of vertebrates
0275Genetically modified vertebrates, e.g. transgenic
C12N 15/10
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
10Processes for the isolation, preparation or purification of DNA or RNA
C12N 15/79
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
C12N 9/0065
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes; Proenzymes; Compositions thereof
0004Oxidoreductases (1.)
0065acting on hydrogen peroxide as acceptor (1.11)
C12P 21/00
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
21Preparation of peptides or proteins
Applicants
  • UNIVERSITY OF MASSACHUSETTS MEDICAL CENTER [US/US]; 55 Lake Avenue North Worcester, MA 01655, US
Inventors
  • LEONARD, Jack, L.; US
  • NEWBURGER, Peter, E.; US
Agents
  • CLARK, Paul, T.; Fish & Richardson P.C. 225 Franklin Street Boston, MA 02110-2804, US
  • FASSE, J., Peter; Fish and Richardson P.C. 225 Franklin Street Boston, MA 02110-2804, US
Priority Data
08/277,49219.07.1994US
08/473,49607.06.1995US
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) POSITIONAL CONTROL OF SELENIUM INSERTION IN POLYPEPTIDES FOR X-RAY CRYSTALLOGRAPHY
(FR) COMMANDE DE POSITION DE L'INSERTION DE SELENIUM DANS DES POLYPEPTIDES POUR LA CRISTALLOGRAPHIE AUX RAYONS X
Abstract
(EN)
The invention provides methods for determining the structure of a polypeptide by a) transfecting a cell with (i) a first nucleic acid encoding the polypeptide, wherein at least one, specific codon of mRNA transcribed from the first nucleic acid is replaced by the codon UGA, and (ii) a second nucleic acid, operably linked to the first nucleic acid, that directs the translation of the UGA codon as selenocysteine only when the cell can obtain selenium from the medium in which it is grown; b) growing the cell in selenium-containing growth medium under conditions in which the cell incorporates at least one selenocysteine residue into the polypeptide at a specific location; c) isolating the polypeptide from the cell or the growth medium; d) forming a crystal of the polypeptide; and e) performing X-ray crystallography on the crystal, wherein the selenocysteine residue is used to determine the structure of the polypeptide.
(FR)
Cette invention se rapporte à des procédés qui permettent de déterminer la structure d'un polypeptide, (a) en transfectant une cellule avec (i) un premier acide nucléique codant ledit polypeptide, dans lequel au moins un codon spécifique d'ARNm transcrit à partir dudit premier acide nucléique est remplacé par le codon UGA, et (ii) un second acide nucléique, lié fonctionnellement au premier acide nucléique, qui dirige la traduction du codon UGA en tant que sélénocystéine, uniquement lorsque la cellule en question peut obtenir du sélénium à partir du milieu dans lequel s'est faite sa croissance; (b) en faisant croître ladite cellule dans un milieu de croissance contenant du sélénium, dans des conditions dans lesquelles la cellule en question incorpore au moins un reste de sélénocystéine dans le polypeptide à un endroit spécifique; (c) en isolant ledit polypeptide de la cellule en question ou de son milieu de croissance; (d) en formant un cristal dudit polypeptide; et (e) en effectuant une cristallographie aux rayons X sur ledit cristal, opération dans laquelle le reste de sélénocystéine est utilisé pour déterminer la structure du polypeptide.
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