tailieunhanh - Molecular cloning and characterization of coat protein gene of banana bract mosaic virus affecting banana cv. Mysore poovan (Aab)

Banana bract mosaic disease (BBrMD) is one of the most important viral diseases of banana which leads to a yield reduction. We have identified banana bract mosaic virus (BBrMV) in banana plants growing in the regions of southern India based on symptomatology, sequence homology, and Serodiagnostic assays. The viral sequence encoding the coat protein was specifically amplified by RT-PCR (Reverse Transcriptase – Polymerase chain reaction) using specific primers bordering the Coat Protein (CP) gene. The unique amplified product thus obtained was cloned into the pGEM-T vector and the authenticity of the cloned gene verified by colony PCR. The nucleotide sequences and the deduced amino acid sequences were compared with the other BBrMV isolates and found to be identical at both the nucleotide and amino acid sequence level of other isolates with 99 to 96 per cent and 95 to 83 per cent respectively. The phylogenetic analysis by the alignment of CP gene sequences of selected 22 isolates also revealed that the present isolate was more similar to KER2 (Kasaragod) isolate. The recombinant clones developed in the present study could be applied in serodiagnosis and genetic engineering. This could be also used as disease diagnostic probes for more sensitive molecular diagnostic techniques like Nucleic acid spot hybridization. | Molecular cloning and characterization of coat protein gene of banana bract mosaic virus affecting banana cv. Mysore poovan (Aab)