Fusion coat protein of pumpkin yellow vein mosaic virus with maltose binding protein: Applications in immunodiagnosis of begomoviruses

南瓜黄脉花叶病毒与麦芽糖结合蛋白的融合外壳蛋白在豇豆花叶病毒免疫诊断中的应用

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作者:Chigurupati Phaneendra, K R S Sambasiva Rao, Reetika Kapoor, R K Jain, Bikash Mandal

Abstract

Availability of adequate quantity of purified virus preparation from plant tissue is the major limitation in producing polyclonal antibodies (PAb) to begomovirus. Very few examples show successful utilization of E. coli expressed recombinant coat protein (CP) for immuno diagnosis of begomoviruses. In the present study, ~771 bp CP gene (~29.0 kDa) of Pumpkin yellow vein mosaic virus (PYVMV) was expressed as a ~71.0 kDa fusion protein with maltose binding protein (MBP) (~42.0 kDa) in E. coli. The MBP-CP was obtained in soluble state. The PAb to the purified fusion protein successfully detected PYVMV and other bipartite and monopartite begomoviruses in the field samples at 1:250 dilution in enzyme linked immunosorbent assay. Our study for the first time showed that MBP-tag fusion CP was suitable to produce diagnostic antibody to begomoviruses.

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