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Chunha Shin,송일찬,이예지,So-Hyeon Baek,김대헌,고기성 한국원예학회 2020 원예과학기술지 Vol.38 No.2
Plant molecular biofarming has been increasingly studied in recent decades. Many kinds ofrecombinant immunotherapeutic proteins have been produced in transgenic tobacco plants. However, tobacco mosaic virus (TMV) can damage tobacco plants and cause pathogenic symptoms,which affects plant biomass production. Cigarette sap solution has been used to infect TMV duringtobacco cultivation. In this study, we obtained TMV-infected transgenic plants expressing a prostatespecificantigen (PSA) fused to an IgG Fc with KDEL ER retention motif (PSA-IgG FcK). Thetypical TMV-associated symptoms appeared and increased on plant leaves 5 days after infection. mRNA expression levels and the presence of TMV were confirmed by RT-PCR and transmissionelectron microscopy analyses. The expression level and purity of the target protein were notsignificantly different between noninfected and infected transgenic plants. TMV was not found inthe purified protein samples from infected plants. Our study showed that TMV pathogen-infectedplant biomass can be harvested and processed to obtain therapeutic vaccine proteins.
Shin Chunha,김기범,Kang Yangjoo,Kim Deuk-Su,Seo Young-Jin,박세라,김미경,Lee Young Koung,김도선,Ko Kisung 한국식물생명공학회 2022 Plant biotechnology reports Vol.16 No.6
Prostate-specifc antigen (PSA) is a protein highly expressed in cancer cells of the prostate gland. In this study, the PSA recombinant protein was fused to the human immunoglobulin G crystallizable fragment (Fc) with the endoplasmic reticulum (ER) Lys-Asp-Glu-Leu (KDEL) retention signal tag for the plant expression system. Agrobacterium-mediated plant transformation was applied to generate transgenic tobacco plants expressing PSA tagged to KDEL (PSAK), PSA-Fc, and PSA-Fc fused to KDEL (PSA-FcK). Western blot analysis showed that the PSA-FcK protein was the most highly expressed among the three diferent proteins in the transgenic plant leaf. The PSAK, PSA-Fc, and PSA- FcK proteins were successfully purifed from plant leaves. Reduced and non-reduced sodium dodecyl sulfate–polyacrylamide gel electrophoresis revealed that the PSA proteins fused to Fc and FcK were dimerized, whereas the PSA itself was not dimerized. N-glycan analysis demonstrated that the N-glycan profle of the PSA-Fc was mainly the structure with α(1,3)-fucose (Fuc) or β(1,2)-xylose (Xyl) glycan residues, whereas the PSA-Fc tagged with the KDEL ER retention signal had mainly an oligomannose-type structure. ELISA and SPR showed that PSA-FcK had a higher binding activity to Fc gamma receptor I than PSA-Fc and human Fc. In addition, PSA-FcK induced anti-PSA, and thus has the potential to be utilized as a prostate cancer vaccine