TRANSIENT EXPRESSION STUDIES OF BmNPV ANTIVIRAL PROTEINS SERINE PROTEASE AND LIPASE IN MULBERRY PLANT
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Date
2018-08-01
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University of Agricultural Sciences, Bangalore
Abstract
Sericulture is an important agrobased cottage industry. Silkworm Bomyx mori L. is being commercially utilized for the production of silk. Silkworm susceptible to fungal, bacterial, viral and protozoan diseases. Among these, Bombyx mori nuclear polyhedrosis virus (BmNPV) causes grasserie, a viral disease in silkworm which is a major disease causing great economic loss to sericulture industry. The present investigation lays main emphasis on transformation of antiviral genes, serine protease and lipase into mulberry leaves using agro infiltration. BmNPV antiviral proteins were cloned in the plant expression vector pBI121. The plant expression vector pBI121 was successfully moved into Agro-bacterium cells for agro infiltration studies in mulberry. The gene integration and expression of antiviral proteins were confirmed by PCR amplification and SDS-PAGE analysis, which was found to be 885 bp of lipase
gene with protein size of 29 Kda, and 855 bp of serine protease gene with protein size of 24 Kda. The antiviral protein genes lipase and serine protease were cloned into bacterial expression vector pET32a for bioassay studies. Bioassay studies were carried out in hybrid silkworm Kolar Gold, by feeding antiviral proteins isolated from bacterial expression system. Silkworms fed with Bombyx mori serine protease and Bombyx mori lipase antiviral proteins against BmNPV infection have showed less mortality when compared with control silkworm with antiviral proteins.