FIG 2.

Parvovirus-derived myosin fusion is expressed in guinea pig. (A) The presence of a parvovirus-derived rep gene in the genome of guinea pig was verified by PCR from genomic DNA. F1/R1 primers were used to amplify regions of the rep gene carried by an EPV (enRep). F2/R2 primers were used to amplified rep in its immediate genomic context (enRepGC). F1/R3 primers were used to amplified rep and Myo9-like exons (enRep-M9l). All amplicons were verified by sequencing. The locations of the primers on the gene are shown as follows: rep hexon in white, Myo9-like exons in different shades of gray, and introns as lines. pb, base pairs. (B) The expression of enRep-M9l transcript was detected in several guinea pig tissues. Total RNA was isolated, and cDNA was generated using oligo(dT) and used for PCR to detect enRep (enRep), enRep-M9l up to the exon 2/3 junction (enRep-M9l), or the full-length enRep-M9l coding sequence (FLenRep-M9l). C. porcellus GAPDH (CpGAPDH) mRNA was used as loading control. The locations of the primers on the enRep-M9l coding sequence are shown under the blots. Exon 5, in which alternative splicing occurs, is indicated by a red asterisk. A DNA migration ladder is indicated on the left-hand side. (C) The enRep-M9l coding sequence confirmed by sequencing of cDNA is shown. The start of each exon is indicated by a bold capital letter. The sequence missing in the alternatively spliced version is shown in red, and the sequence of enRep is underlined. (D) Alignment of the long (enRep-M9lL) and short (enRep-M9lS) proteins encoded by the cloned sequences. The Rep domain is framed in red.