Methods for improving recombinant protein expression
Materials and methods are provided which allowed for increased expression of a transfected gene of interest in a recombinant host cell.
1. A method for increasing heterologous protein expression in a mammalian host cell comprising the step of culturing the host cell under conditions that allow for protein expression, said host cell comprising a first heterologous polynucleotide sequence encoding said heterologous protein, said host cell further comprising a second polynucleotide sequence having a protein coding sequence for a selectable marker protein, said second polynucleotide having a sequence modification compared to a wild-type polynucleotide encoding said selectable marker protein, said sequence modification reducing translation efficiency of mRNA encoded by said second polynucleotide, said second polynucleotide having said sequence modification and said wild-type polynucleotide encoding identical amino acid sequences for said selectable marker protein, said first polynucleotide and said second polynucleotide encoded on a Chinese hamster elongation factor 1 (CHEF1) expression vector, wherein the second polynucleotide comprises the polynucleotide sequence of SEQ ID NO: 3 or SEQ ID NO: 4.