Transient expression of the full‐length glycoprotein from infectious hematopoietic necrosis virus in bean (Phaseolus vulgaris) leaves via agroinfiltration

Abstract

The glycoprotein of infectious hematopoietic necrosis virus (IHNV), the causative agent of acute disease in salmonids, is the only structural protein of the virus that can induce protective immunity in the fish host. Here, the reliability of bean (Phaseolus vulgaris) plant for the production of this viral protein was examined by the transient expression method. Using the syringe agroinfiltration method, leaves of bean plants were transformed with the expression construct encoding the full-length of IHNV glycoprotein (IHNV-G) gene. Furthermore, the transformation efficacy of two infiltration buffers including PBS-A (PBS+acetosyringone) and MMS-A (MES buffer + MgSO 4 + sucrose + acetosyringone) was compared. The analysis of mRNA and dot-blot assay confirmed the transcription and translation of IHNV-G protein in bean leaves. Moreover, Western blotting verified the production of intact, full-length (∼57 kDa) IHNV-G protein in the agroinfiltrated plants. Of note, the production level of IHNV-G using MMS-A agroinfiltration buffer was approximately five times higher compared to PBS-A buffer (0.48 vs. 0.1% of total soluble protein), indicating the effect of infiltration buffer on the transient transformation efficiency. The recombinant protein was purified at the final yield of 0.35 μg/g of fresh leaf tissue, using nickel affinity chromatography. The present work is the first report describing the feasibility of the plant expression platform for the production of IHNV-G protein, which can be served as an oral vaccine against IHNV infection.

Molecular farming: Expanding the field of edible vaccines for sustainable fish aquaculture

“…Recently, Fazeli et al 224 described the production strategy of recombinant glycoprotein as the only immunostimulant protein of the 228 produced bacterial protein p57 from Renibacterium salmoninarum using microalga Ch. reinhardtii as the production platform.…”

“…Therefore, attempts to use legumes as a production platform for fish vaccination are warranted. Recently, Fazeli et al 224 described the production strategy of recombinant glycoprotein as the only immunostimulant protein of the infectious haematopoietic necrosis virus infecting salmonids. After agroinfiltration of bean leaves ( Phaseolus vulgaris ), recombinant glycoprotein production has reached 0.48% of total soluble protein.…”

Comparative Evaluation of Transient Protein Expression Efficiency in Tissues across Soybean Varieties Using the Tsukuba System

“…A transient expression by agroinfiltration has been demonstrated in some legume species, including the common bean ( Phaseolus vulgaris ) [ 9 , 10 ], Medicago truncatula [ 11 ], cowpea ( Vigna unguiculata ) [ 12 ], lotus ( Lotus japonicus ) [ 10 , 13 ], pea ( Pisum sativum ) [ 14 , 15 ], and Caragana intermedia [ 16 ]. Transient protein expression has been reported in soybean pods [ 10 ], protoplasts [ 17 ], and roots [ 18 ].…”