Detection of pepper leaf curl virus through PCR amplification and expression of its coat protein in Escherichia coli for antiserum production

Abstract

Pepper leaf curl virus (PepLCV) is the most destructive pathogen of pepper and causes substantial economic losses of chilli production worldwide. Curling and puckering of leaves and stunted growth of the plants are typical symptoms of the viral infection. For a reliable detection of PepLCV, coat protein specific primer pairs were designed and after polymerase chain reaction amplification (PCR), -1 kb amplification product specific to coat protein gene was amplified. Cloning of this amplification product in an expression vector resulted in high-level expression of a 28 kD protein. Coat protein gene product is important for vector specificity and is also responsible for viral capsid formation and whiteflymediated transmission. The purified protein can be used for production of coat protein specific antisera leading to development of an efficient detection system for this very important viral pathogen of pepper.

Key words: PepLCV, geminivirus, virus detection, polymerase chain reaction amplification (PCR), immunoblot.