Patent ID: 6610472
Filing Date: 2003-08-26
Classification: C07K,C12Q,C40B,G01N

Abstract:
A method for generating a library of yeast expression vectors encoding a library of protein complexes, comprising:transforming into yeast cells a library of insert nudeotide sequences that are linear and double-stranded, and a library of linearized yeast expression vectors, each having a 5â€²- and 3â€²-terminus sequence at the site of linearization; and having homologous recombination occur between the vector and the insert sequence such that the insert sequence is included in the vector in the transformed yeast cells, wherein each of the linearized yeast expression vectors in the vector library comprises a first polynucleotide sequence encoding a first polypeptide subunit which varies within the vector library and has a diversity of at least 1Ã—103; the insert sequences of the insert library comprise a second nucleotide sequence encoding a second polypeptide subunit which varies within the insert library, each of the insert sequences comprising a 5â€²- and 3â€²-flanking sequence at the respective ends of the insert sequence and being sufficiently homologous to the 5â€²- and 3â€²-terminus sequences of the linearized yeast expression vector, respectively, to enable homologous recombination to occur, and the first and second polypeptide subunits are capable of being expressed as separate proteins and assembling to form the library of protein complexes of the first and second polypeptide subunits and having a diversity of at least 1Ã—106.