INTRON-BASED UNIVERSAL CLONING METHODS AND COMPOSITIONS

The present invention relates to a method for producing a fusion polynucleotide encoding a polypeptide of interest. The method comprises the steps of providing a first polynucleotide and a second polynucleotide, and contacting said first polynucleotide and second polynucleotide with a type IIs restr...

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Hauptverfasser: SCHNEIDER, Marion, LEUSCHNER, Wulf, DITTRICH, Werner, BIRKENFELD, Joerg
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creator SCHNEIDER, Marion
LEUSCHNER, Wulf
DITTRICH, Werner
BIRKENFELD, Joerg
description The present invention relates to a method for producing a fusion polynucleotide encoding a polypeptide of interest. The method comprises the steps of providing a first polynucleotide and a second polynucleotide, and contacting said first polynucleotide and second polynucleotide with a type IIs restriction endonuclease and a ligase under conditions that allow for the cleavage of the first polynucleotide and second polynucleotide by said type IIs restriction endonuclease and the ligation of the resulting cleavage products, thereby producing the fusion polynucleotide encoding the polypeptide of interest. The first polynucleotide comprises the 5′ portion of an intron, and the second polynucleotide comprises the 3′ portion of an intron. Further envisaged by the present invention is a polynucleotide encoding a polypeptide of interest, which, when transcribed in a eukaryotic host cell, is transcribed into a transcript comprising at least five introns which are heterologous to said polynucleotide.
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The method comprises the steps of providing a first polynucleotide and a second polynucleotide, and contacting said first polynucleotide and second polynucleotide with a type IIs restriction endonuclease and a ligase under conditions that allow for the cleavage of the first polynucleotide and second polynucleotide by said type IIs restriction endonuclease and the ligation of the resulting cleavage products, thereby producing the fusion polynucleotide encoding the polypeptide of interest. The first polynucleotide comprises the 5′ portion of an intron, and the second polynucleotide comprises the 3′ portion of an intron. 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subjects BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
title INTRON-BASED UNIVERSAL CLONING METHODS AND COMPOSITIONS
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