PRODUCTION OF POLYCLONAL ANTIBODIES SPECIFIC TO CITRUS TRISTEZA VIRUS (CTV) USING THE 6X HIS-TAGGED FUSION COAT PROTEIN

Abstract

Citrus tristeza virus (CTV) is one of the most destructive and economically
important diseases of commercial citrus worldwide. In this study, the coat protein
(cp) gene of CTV was subcloned into PQE-30 vector at Bam HI and Hind-III
restriction sites. The constructed plasmid that called pRMAF2 was transformed into
Escherichia coli M15 strain for its expression and production of fusion protein for
CTV-cp gene. The suitable condition for inductions of CTV-cp was in the presence
of 1 mM IPTG and overnight incubation time. The bacterial culture was harvested
and the expressed fusion protein(s) was purified by QIA express kit. Polyclonal
antibodies (PAbs) were raised by injection of the purified fusion protein(s) into two
white mice followed by western blotting analysis using the expressed fusion
protein(s) and the CTV-infected citrus extracts as controls. It can be concluded that
the immuno-enzymatic detection based on the produced antibodies against CTV-cp-
ORF fusion proteins were safe, speed and sensitive for the detection of the virus