title: Expression of a Human Prostatic Acid Phosphatase (PAP)-IgM Fc Fusion Protein in Plants UsingIn vitroTissue Subculture.
abstract: In this study, prostatic acid phosphatase (PAP), which is overexpressed in human prostate cancer cells, was cloned to be fused to the IgM constant fragment (Fc) for enhancing immunogenicity and expressed in transgenic tobacco plants. Then, the transgenic plants were propagated byin vitrotissue subculture. Gene insertion and expression of the recombinant PAP-IgM Fc fusion protein were confirmed in each tested the first, second, and third subculture generations (SG1, SG2, and SG3, respectively). Transcription levels were constantly maintained in the SG1,SG2, and SG3leaf section (top, middle, and base). The presence of the PAP-IgM Fc gene was also confirmed in each leaf section in all tested subculture generations. RNA expression was confirmed in all subculture generations using real-time PCR and quantitative real-time PCR. PAP-IgM Fc protein expression was confirmed in all leaves of the SG1, SG2, and SG3recombinant transgenic plants by using quantitative western blotting and chemiluminescence immunoassays. These results demonstrate that the recombinant protein was stably expressed for several generations ofin vitrosubculture. Therefore, transgenic plants can be propagated usingin vitrotissue subculture for the production of recombinant proteins.
authors.affiliation: Therapeutic Protein Engineering Laboratory, Department of Medicine, College of Medicine, Chung-Ang University Seoul, South Korea.
authors.forename: Yang J
authors.initials: YJ
authors.lastname: Kang
authors.affiliation: Therapeutic Protein Engineering Laboratory, Department of Medicine, College of Medicine, Chung-Ang University Seoul, South Korea.
authors.forename: Deuk-Su
authors.initials: DS
authors.lastname: Kim
authors.affiliation: Department of Urology, College of Medicine, Chung-Ang University Seoul, South Korea.
authors.forename: Soon-Chul
authors.initials: SC
authors.lastname: Myung
authors.affiliation: Therapeutic Protein Engineering Laboratory, Department of Medicine, College of Medicine, Chung-Ang University Seoul, South Korea.
authors.forename: Kisung
authors.initials: K
authors.lastname: Ko
keywords: propagation; prostatic acid phosphatase; recombinant protein; subculture generation; transgenic plant