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Figure 1 | Virology Journal

Figure 1

From: A baculovirus-mediated strategy for full-length plant virus coat protein expression and purification

Figure 1

Gene and protein schemes with deduced amino acid sequence. (A) The polh-6xhis fragment was amplified and cloned into the commercial vector (I), pFB1 to generate pFB1-polh-6xhis (not shown). We used BglII (primer added) and NotI (from the pGem-T® easy vector) restriction sites to clone the modified gene and NcoI (primer added) restriction site to use for virus coat protein fusion (II and III). These vectors were used to construct recombinant viruses, vAc-polh-6xhis and vAc-GarMbFV-cp-polh-6xhis by site-specific transposition in E. coli (Bac-to-bac® system, Invitrogen). The virus expressing non-fused GarMbFV-CP was constructed by homologous recombination inside insect cells co-transfected with DNA from pSyn-GarMbFV-cp and vSynVI-gal (see Methods). Deduced amino acid sequence of the (B) non-fused coat protein, GarMbFV-CP (27.9 kDa), (C) Polh-6xHis (29.9 kDa), and (D) Polh-GarMbFV-CP-6xHis recombinant protein (50.0 kDa) are shown.

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