Extending the Serum Half-Life of G-CSF via Fusion with the Domain III of Human Serum Albumin

Figure 1

Construction of recombinant pPIC-3DHSA-G-CSF eukaryotic expression vector. The synthesized HSA and G-CSF genes were used as templates for the amplification of 3DHSA-G10 and H10-G-CSF genes by PCR with primer 1 and primer 2, primer 3 and primer 4, respectively. The purified 3DHSA-G10 and H10-G-CSF genes were used as templates for obtaining 3DHSA-G-CSF gene by overlap PCR. Both pPICZαA plasmid and 3DHSA-G-CSF gene were digested by XhoI and ligated together by T4 DNA ligase to construct expression vector pPIC-3DHSA-G-CSF. 3DHSA-G-CSF was inserted into the sites XhoI of pPICZαA along with the open reading frame of the α-factor signal under the control of the AOX1 promoter. The KEX2 cleavage site was located between the signal peptide and the 3DHSA-G-CSF sequences.