Skip to main content
. 2007 Sep 5;81(22):12156–12168. doi: 10.1128/JVI.01591-07

FIG. 2.

FIG. 2.

One-base mismatch extension by RIO vector. (A) Experimental scheme. (B) 30TM acceptor. Newly synthesized DNA transfers from a donor template to an acceptor template with 30 bases of identity. If the DNA transfers at any position within the 30 bases of identity on the donor template, an AscI site will be generated in the provirus. (C) 1TMM acceptor. The 1TMM acceptor contains 29 bases of identity and 1 mismatched base. If DNA transfers from the 5′ end of the donor and the mismatch extends, an AscI site will be generated. If the DNA transfer occurs prior to the template end and no mismatch extension occurs, the template base will be incorporated and no AscI site will be formed. Acceptor region abbreviations are given in the legend to Fig. 1. (D) PCR amplification of puromycin resistance gene from pooled proviral DNA by use of primers indicated in Fig. 1C and analysis with AscI digestion. pRSVpuro, a vector containing an intronless puromycin resistance gene amplified with primers SF304 and MK204. RIO vectors, with their artificial introns, yield a longer amplification product. PCR products from 30TM and 1TMM, undigested and digested with AscI, are shown. Alternate targets that appear in the 1TMM product pool are indicated. puro, puromycin.