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Expression of recombinant Apple chlorotic leaf spot virus coat protein in heterologous system: production and use in immunodiagnosis
Authors:Tanuja Rana  Vanita Chandel  Vipin Hallan  Aijaz A Zaidi
Institution:1. Plant Virology Laboratory, Floriculture Division, Institute of Himalayan Bioresource Technology (CSIR), Palampur, 176 061, Himachal Pradesh, India
Abstract:Expression condition for maximum recovery of recombinant Apple chlorotic leaf spot virus (ACLSV) coat protein was standardized. The in vitro expressed fusion protein with 6xHis tag (~43 Kd) was purified from inclusion bodies and used as an antigen for raising polyclonal antiserum in rabbit. This antiserum consistently detected ACLSV in pome and stone fruits as well as herbaceous host plants by direct double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA) and direct tissue blot immunoassay (DTBIA). The conditions for immuno-capture RT-PCR (IC-RT-PCR) were also standardized.
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