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

摘要

In order to devise a method for exploring a CHO cell hotspot region for high expression of a recombinant gene, a CHO cell line which integrated a vector expressing enhanced green fluorescent protein (eGFP) in the genome was constructed, the CHO cell line highly expressing GFP was firstly isolated through a flow cytometer equipped with an automatic cell deposition device is firstly separated and secondarily separated by obtaining mono clone through a limiting dilution method, and a target PCR was used to amplify an insertion site of the eGFP gene to obtain the sequence of the gene insertion site through next-generation sequencing, thereby finding the hot spot region through sequence analysis.