Structure of PAP-IgM FcK fusion protein with J-chain expressed in transgenic plant

Abstract

Transgenic plants expressing immunoglobulin (Ig) M Fc-fused Prostate acid phosphatase (PAP) antigenic proteins (PAP-IgM FcK) and J-chain proteins were generated by Agrobacterium-mediated transformation. The Fc region was tagged with the ER retention motif (KDEL) to make PAP-IgM FcK. Two transgenic plants were crossed together to generate F1 expressing both PAP-IgM FcK and J-chain proteins (PAP-IgM FcK × J-chain). PCR and RT-PCR analyses confirmed the transgene insertion and mRNA transcription of PAP-IgM FcK and J-chain in leaf tissue of PAP-IgM FcK × J-chain F1 plant. Western blot confirmed the expression of PAP-IgM FcK × J-chain protein. Size exclusion (SEC)-high performance liquid chromatography (HPLC) and Bio-transmission electron microscope (TEM) analyses were performed to show the size and shape of the PAP- IgM FcK × J-chain fusion proteins. These results suggest that PAP-IgM FcK with J-chain can be produced in plant expression system with plant crossing.

Keywords

• transgene
• plant biotechnology
• immunoglobulin
• protein engineering