J链PAP-IgM-FcK融合蛋白在转基因植物中表达的结构

IF 1.2 Q3 MULTIDISCIPLINARY SCIENCES

The EuroBiotech Journal Pub Date : 2023-01-01 DOI:10.2478/ebtj-2023-0006

Y. Kang, Deuk‐Su Kim, Kibum Kim, S. Myung, Y. J. Oh, Sungsu Park, P. Hinterdorfer, K. Ko

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引用次数: 2

摘要

摘要通过农杆菌介导的转化，获得了表达免疫球蛋白（Ig）M-Fc融合的前列腺酸性磷酸酶（PAP）抗原蛋白（PAP-IgM-FcK）和J链蛋白的转基因植物。Fc区域用ER保留基序（KDEL）标记以制备PAP-IgM-FcK。将两个转基因植物杂交在一起以产生表达PAP-IgM-FcK和J链蛋白的F1（PAP-IgM-FcK×J链）。PCR和RT-PCR分析证实了PAP-IgM-FcK和J链在F1植株叶片组织中的转基因插入和mRNA转录。蛋白质印迹证实PAP-IgM-FcK×J链蛋白的表达。通过大小排阻（SEC）-高效液相色谱（HPLC）和生物透射电子显微镜（TEM）分析显示了PAP-IgM-FcK×J链融合蛋白的大小和形状。这些结果表明，具有J链的PAP-IgM-FcK可以在具有植物杂交的植物表达系统中产生。

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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.

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来源期刊

The EuroBiotech Journal Agricultural and Biological Sciences-Food Science

CiteScore

3.60

自引率

0.00%

发文量

审稿时长

10 weeks