Directional cloning of Citrus psorosis coat protein gene and protein expression

The full length of the coat protein gene of two Turkish isolates of Citrus psorosis virus was amplified by RT-PCR. The product of the CP-gene was cloned into the TA vector for sequencing in both directions, sense, and antisense. The full length of ORF CP-gene (1318 bp) of CPsV was cloned into pET101/D-TOPO with blunt end ligation. The 20 positive colonies for CPsV-CP gene were checked by PCR and sequenced for both ends. The correct direction insertion of the colonies was identified and a few of them were selected for protein expression. The Western blot was run for the CPsV-CP gene expression. The 48.6 kDa protein was expressed by induced cells with IPTG including CPsV-CP in 4 selected bacterial cells. The expressed coat protein of CPsV was shown with western blotting