Enhanced cell-free protein expression by fusion with immunoglobulin Ckappa domain.
While cell-free systems are increasingly used for protein expression in structural and functional studies, several proteins are difficult to express or expressed only at low levels in cell-free lysates. Here, we report that fusion of the human immunoglobulin kappa light chain constant domain (Ckappa) at the C terminus of four representative proteins dramatically improved their production in the Escherichia coli S30 system, suggesting that enhancement of cell-free protein expression by Ckappa fusion will be widely applicable.
Palmer
Elizabeth
E
Technology Research Group, The Babraham Institute, Cambridge CB2 4AT, United Kingdom.
Liu
Hong
H
Khan
Farid
F
Taussig
Michael J
MJ
He
Mingyue
M
D002474:Cell-Free System / Q000502:physiology; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis*
D007145:Immunoglobulin kappa-Chains; D011993:Recombinant Fusion Proteins