Article
A simple novel method for the preparation of noncovalent homodimeric, biologically active human interleukin 10 in Escherichia coli-enhancing protein expression by degenerate PCR of 5' DNA in the open reading frame.
Protein expression and purification - 1 Dec 2008
Klompus Shelley, Solomon Gila, Gertler Arieh
Abstract excerpt
DNA inserts encoding human interleukin 10 (hIL-10), optimized for codon usage and secondary RNA structure, were purchased from several commercial sources and subcloned into a pMon vector. Despite the optimization, protein expression was nil. We therefore subjected the 5' segment of the cDNA encoding N-terminal amino acids 2-11 to degenerate PCR in order to create a small library of 130K theoretical cDNA...
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