Figure 1 : Schematic representation of the construction and production of Pfs25 and Pfs25-IMX313 in viral vectors and P. pastoris.

From: Enhancing immunogenicity and transmission-blocking activity of malaria vaccines by fusing Pfs25 to IMX313 multimerization technology

Figure 1

(A) Four constructs used in these experiments: in viral vectors 1) Pfs25 (aa 22–194) fused to an N-terminal secretion signal peptide tPA; 2) Pfs25-IMX313 fused to tPA; and in P. pastoris 3) Pfs25 (aa 22–194) with a C-terminal hexahistidine (His6) tag; 4) Pfs25 fused to IMX313 with an N-terminal His6 tag. (B) Schematics showing the steps in production of Pfs25 and Pfs25-IMX313 in P. pastoris. The antigen sequences were cloned into the P. pastoris expression plasmid pPinkα-HC (InvitrogenTM, Life Technologies, UK) (containing the α-mating factor secretion signal peptide). After electroporation of target plasmids into P. pastoris, positive colonies were screened for protein expression and successful candidates were selected for protein production as indicated. (C) Idealised structure of the Pfs25-IMX313 heptamer. Pfs25 (blue) homology model based on the crystal structure of Pvs25 (PDB: 1Z27) fused N-terminally to IMX313 represented here by the crystal structure of the human C4bp (PDB: 4B0F).