Effect of the hydrophobin HFBI-fusion tag on exogenous protein accumulation in tobacco plant
10.3969/j.issn.1673-4254.2015.12.01
- VernacularTitle:疏水蛋白HFBI融合标签能显著提高外源基因在植物中的表达水平
- Author:
Xiqian ZHANG
1
;
Hongzhen MU
;
Ting MA
;
Xiangzhen DING
;
Zhiying LI
;
Sheng WANG
Author Information
1. 西部特色生物资源保护与利用教育部重点实验室
- Keywords:
hydrophobin;
fusion tags;
green fluorescent protein;
Trichoderma reesei;
Nicotiana benthamiana
- From:
Journal of Southern Medical University
2015;(12):1665-1671
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the mechanisms by which HFBI fusions increase recombinant fusion protein accumulation in plants. Methods The HFBI sequence from Trichoderma reesei was synthesized and two plant expression vectors for expression of green fluorescence protein (GFP) and GFP-HFBI were constructed. The vectors were inoculated in Nicotiana benthamiana plants through agroinfiltration, and the expression levels and mRNA accumulation levels of GFP in Nicotiana leaves were examined by Western blotting, ELISA and RT-PCR. Results The HFBI fusion tag significantly enhanced the accumulation of GFP in the leaves of N. benthamiana without causing toxic effects. Endoplasmic reticulum-targeted GFP-HFBI fusion induced the formation of spherical protein particles in the plant cells. Conclusion HFBI fusions can increase the accumulation of its fusion partner in plants by forming stable protein particles, which probably shields the target protein from endogenous protease-induced degadation. HFBI fusion technology provides an alternative to improving recombinant protein expression in plants from agroinfection-compatible expression vectors.
