Optimization of the human colorectal carcinoma antigen GA733-2 production in tobacco plants

被引：4

作者：

Park, Se Hee ^{[1
]}

Ji, Kon-Young ^{[2
]}

Kim, Hyun Min ^{[1
]}

Ma, Sang Hoon ^{[1
]}

Park, Seo Young ^{[1
]}

Do, Ju Hui ^{[1
]}

Oh, Doo-Byoung ^{[3
,4
]}

Kang, Hyung Sik ^{[1
]}

Shim, Jae Sung ^{[1
]}

Joung, Young Hee ^{[1
]}

机构：

[1] Chonnam Natl Univ, Sch Biol Sci & Technol, Gwangju 61186, South Korea

[2] Korea Inst Oriental Med, Herbal Med Res Div, Daejeon 34054, South Korea

[3] Korea Res Inst Biosci & Biotechnol KRIBB, Synthet Biol & Bioengn Res Ctr, Daejeon 34141, South Korea

[4] Univ Sci & Technol UST, Dept Biosyst & Bioengn, Daejeon 34113, South Korea

来源：

PLANT BIOTECHNOLOGY REPORTS | 2021年 / 15卷 / 01期

关键词：

Colorectal cancer; GA733-2; Plant-derived therapeutic protein; Transgenic plants; Transient expression;

D O I：

10.1007/s11816-020-00657-y

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

The colorectal carcinoma-associated protein GA733-2 is one of the representative candidate protein for the development of plant-derived colorectal cancer vaccine. Despite of its significant importance for colorectal vaccine development, low efficiency of GA733-2 production limits its wide applications. To improve productivity of GA733-2 in plants, we here tested multiple factors that affect expression of recombinant GA733-2 (rGA733-2) and rGA733 fused to fragment crystallizable (Fc) domain (rGA733-Fc) protein. The rGA733-2 and rGA733-Fc proteins were highly expressed when the pBINPLUS vector system was used for transient expression in tobacco plants. In addition, the length of interval between rGA733-2 and left border of T-DNA affected the expression of rGA733 protein. Transient expression analysis using various combinations of Agrobacterium tumefaciens strains (C58C1, LBA4404, and GV3101) and tobacco species (Nicotiana tabacum cv. Xanthi nc and Nicotiana benthamiana) revealed that higher accumulation of rGA733-2 and rGA733-Fc proteins were obtained by combination of A. tumefaciens LBA4404 and Nicotiana benthamiana. Transgenic plants generated by introduction of the rGA733-2 and rGA733-Fc expression cassettes also significantly accumulated corresponding recombinant proteins. Bioactivity and stability of the plant-derived rGA733 and rGA733-Fc were evaluated by further in vitro assay, western blot and N-glycosylation analysis. Collectively, we here suggest the optimal condition for efficient production of functional rGA733-2 protein in tobacco system.

引用

页码：55 / 67

页数：13