Transformation of eukaryotic cells using two different recombination sites recognised by two different unidirectional site-specific recombinases

摘要

A site-specifically integrating target vector that comprises: (a) A first vector recombination site that recombines with a genomic recombination site in the presence of a first unidirectional site-specific recombinase; (b) A second vector recombination site that recombines with donor recombination site in the presence of a second unidirectional site-specific recombinase that is different from the first unidirectional site-specific recombinase; (c) A promoter-less first portion of first selectable marker adjacent to the second vector site's 3' end; and (d) A second selectable marker that is different from the first selectable marker. The vector is used in conjunction with a donor vector that comprises a polynucleotide encoding a protein of interest, a donor recombination site and a second portion of a first selectable marker which comprises a promoter. The vector is used in methods for integrating a protein of interest into the genome of a eukaryotic cell for protein production.