Fig. 5.

Confirmations of gag insert deletion during in vivo replication of the rCDV-SIVgag1 in ferret tissues by traditional RT-PCR and sequencing. (A) RNA extracted from ferret tissues was RT-PCR amplified and run on a 1% agarose gel. Lane 1, TrackIt™ 1 kb Plus DNA Ladder; lane 2, mesenteric lymph nodes 7 days after intramuscular injection; lane 3, mesenteric lymph nodes 7 days after intranasal inoculation; lane 4, large intestines high density Peyer's patches 28 days after intramuscular injection; lane 5, RNA positive control; lane 6, plasmid DNA positive control; lane 7, no template negative control. (B) The solid line indicates the 1.6 kb gag mRNA with the solid box showing the amplicon used in the SIV gag specific gRNA RT-qPCR (see Fig. 1D). The dotted line indicates the position of the 1416 Nt deletion as determined by sequencing.