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ORF cDNA Clones/MolecularCloud> | Virus detection was done by RT-PCR using total RNA as described by Halgren et al. [11]. RT was performed using random hexamers and RevertAid® reverse transcriptase (Thermo Scientific, USA) following manufacturer’s instructions. PCR was carried out in a 10 μL mixture containing 1 μL 10× buffer, 0.2 uL of 10 mM dNTPs, 1.5 μL cDNA template, 6.8 uL of molecular biology grade water, 0.1 μL of Taq DNA polymerase (GenScript, USA) and 0.2 μL of each primer (40 μM) designed and selected based on specificity across closest relatives. PCR parameters were as follows: 94°C for 4 min, 40 cycles of 94°C for 45 s, 57°C for 30 s and 72°C for 45 s, and a final extension step of 10 min at 72°C. | Get A Quote |
A new member of the genus Potexvirus was fully sequenced and characterized. The virus was isolated from babaco (Vasconcellea x heilbornii), a natural hybrid native to Ecuador. The virus contains a 6,692 nt long genome organized in five open reading frames in an arrangement typical of other potexviruses. Sequence comparisons revealed close relatedness with Papaya mosaic virus (PapMV), Alternathera mosaic virus (AltMV) and Senna mosaic virus (SenMV), exhibiting nucleotide identities up to 67% for the polymerase (Pol) and 68% for the coat protein (CP), with deduced amino acid identities of 70% and 72% for the Pol and CP, respectively. The presence of an AlkB domain, in the polymerase region, was observed. ... More