Abstract
Sweet potato leaf curl virus (SPLCV) causes yield losses in sweet potato cultivation. Diagnostic techniques such as serological detection have been developed because these plant viruses are difficult to treat. Serological assays have been used extensively with recombinant antibodies such as whole immunoglobulin or single-chain variable fragments (scFv). An scFv consists of variable heavy (VH) and variable light (VL) chains joined with a short, flexible peptide linker. An scFv can serve as a diagnostic application using various combinations of variable chains. Two SPLCV-specific scFv clones, F7 and G7, were screened by bio-panning process with a yeast cell which expressed coat protein (CP) of SPLCV. The scFv genes were subcloned and expressed in Escherichia coli. The binding affinity and characteristics of the expressed proteins were confirmed by enzyme-linked immunosorbent assay using SPLCV-infected plant leaves. Virus-specific scFv selection by a combination of yeast-surface display and scFv-phage display can be applied to detection of any virus.
Highlights
Sweet potato leaf curl virus (SPLCV) causes yield losses in sweet potato cultivation
V1 protein of SPLCV Haenam 1 strain was amplified by polymerase chain reaction (PCR) and the products were visualized in the form of a slight single band product on 1% agarose gel containing ethidium bromide (Fig. 2A)
The amplified DNA fragments from the V1 of SPLCV was cloned into a pCTCON plasmid vector for yeast-surface display
Summary
Sweet potato leaf curl virus (SPLCV) causes yield losses in sweet potato cultivation. Diagnostic techniques such as serological detection have been developed because these plant viruses are difficult to treat. Serological assays have been used extensively with recombinant antibodies such as whole immunoglobulin or single-chain variable fragments (scFv). Two SPLCV-specific scFv clones, F7 and G7, were screened by bio-panning process with a yeast cell which expressed coat protein (CP) of SPLCV. The sweet potato leaf curl virus (SPLCV), a member of the genus Begomovirus, is transmitted by the whitefly (Bemisia tabaci Genn.), which is the only natural vector[8]. Several detection methods employing polymerase chain reaction (PCR) of SPLCV genomic DNA have been reported[11,12,13]. Animals or animal cell cultures are required for plant virus diagnosis in plant virus laboratories
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