Patent ID: 6291160
Filing Date: 2001-09-18
Classification: A61K,C07K

Abstract:
A method of producing a diverse population of vectors comprising a first polynucleotide sequence encoding a first protein and a second polynucleotide sequence encoding a second protein said first and second proteins being capable of forming a heterodimeric receptor, wherein said first polynucleotide sequences and said second polynucleotide sequences have a translational orientation, said method comprising the steps,(a) synthesizing a first genetic library containing a plurality of diverse first polynucleotide sequences, wherein said library comprises first cloning vectors, each vector in said library comprising a single first polynucleotide sequence, and said first polynucleotide sequence is adjacent to a first restriction endonuclease recognition site, wherein said first restriction endonuclease recognition site is located proximal upstream to the translation initiation site of said first polynucleotide sequence;(b) synthesizing a second genetic library containing a plurality of diverse second polynucleotide sequences, wherein said library comprises second cloning vectors, each vector in said library comprising a single second polynucleotide sequence, and said second polynucleotide sequence is adjacent to a second restriction endonuclease recognition sites, wherein said second restriction endonuclease recognition site is located proximal upstream to the translation initiation site of said second polynucleotide sequence, wherein the first restriction endonuclease recognition site and the second restriction endonuclease recognition site produce ligation compatible ends upon digestion;(c) digesting said first and second genetic libraries with restriction endonucleases capable of specifically cleaving said first and second restriction endonuclease recognition sites, whereby first and second digested libraries are produced; and(d) ligating said first and second digested libraries via their ligation compatible ends.