Sanofi
INTRON-BASED UNIVERSAL CLONING METHODS AND COMPOSITIONS

Abstract:

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.