Exploring hotspot method for expression of recombinant protein in cell line using next-generation sequencing

摘要

The present inventors constructed a CHO cell line integrated with a vector expressing enhanced green fluorescent protein (eGFP) in the genome to construct a method for searching CHO cell hot spot region for high expression of a recombinant gene, Highly expressing CHO cell line is firstly isolated by flow cytometry equipped with automatic cell deposition device, secondly separated by monoclonal acquisition by limiting dilution method, amplified the insertion position of eGFP gene through target PCR, and sequenced by next generation The sequence of the gene insertion site was obtained and the hot spot region was found through sequence analysis.