Patent ID: 9133485
Filing Date: 2015-09-15
Classification: C12N

Abstract:
1. A method for the generation of a high producer host cell for the production of recombinant gene products comprising the following steps: (a) providing a target vector comprising: (b) introducing the target vector in a host cell under conditions in order to allow random integration of the target vector into the genome of the host cell, (c) selecting a host cell having stably integrated the target vector and showing high transcriptional activity of the reporter gene (RG1), (d) providing an exchange vector comprising: (e) introducing the exchange vector into a host cell obtained in step (c) under conditions to allow a double strand break at the first and/or second recognition site as defined in step (a)(iv) and an integration of the exchange vector into the genome of the host cell by double-strand break-mediated homologous recombination, whereby the inactive second selection marker gene (ΔSM2) is activated by an integration of the exchange vector by homologous recombination with the target vector, (f) selecting a high producer cell having integrated the exchange vector by homologous recombination with the integrated target vector, wherein the producer cell expresses the GOI.