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Design of fusion proteins

IG Iram Gull
MA Muhammad Shahbaz Aslam
IT Imran Tipu
RM Roohi Mushtaq
TA Tehseen Zamir Ali
MA Muhammad Amin Athar
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To develop latent interferon alpha 2b (IFNα2b) fusion protein, the latency associated protein (LAP) domain of human TGFβ has been fused in two conformations: i) LAP was fused either at N-terminus or ii) at C- terminus of IFNα2b. At splicing junction of each fusion gene, HCV NS3 protease cleavage site (EDVVCCSMSY) was introduced as cleavable linker. The fusion proteins were designated as LAP-NS3- IFN and IFN-NS3-LAP. The N- terminus of each fusion gene was tagged with peptide sequence “N-Kex2 site-His tag- Gly/Ser spacer-enterokinase site-C” to facilitate processing of protein in pichia pastoris expression system, protein purification and removal of His-tag from protein after purification by enterokinase respectively.

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