The Genomic 5′ Terminus of Manchester Calicivirus

Abstract

An enteric calicivirus showing the classic cup-shaped surface morphology was identified in a stool sample obtained from a child with symptoms of acute gastroenteritis (Portishead virus, PHV). Genomic RNA was extracted directly from the PHV stool sample and amplified by RT-PCR using primers based on the Manchester isolate of HuCV. The 3′ terminus of the cDNA was defined by homopolymer tailing with dATP and revealed an additional 165 nucleotides suggesting that the previously determined Manchester HuCV (MV) genome sequence was incomplete. Homopolymer tailing of MV cDNA primed using sequence data from the 5′ terminus of PHV allowed extension of the MV genome by a further 165 nucleotides thereby increasing the overall genome length to 7431 nucleotides and resulting in an additional 72 amino acids at the N-terminus of the polyprotein. A conserved sequence motif typical of other caliciviruses was also identified at the extreme 5′-terminus of the genome.