FIG. 3.

Confirmation of the presence of ACH.p12^I and ACH.p30^II mutations. (A) RT-PCR with total RNA from ACH.wt- and ACH.p12^I-immortalized cells and primers 5094 and 7135 amplified a 2,041-bp fragment from the gag-pol and env transcripts, as well as a 490-bp pX-I transcript only in the ACH.wt-immortalized cells. Primers specific for human hypoxanthine-guanine phosphoribosyltransferase (HGPRT) were used as an additional control for RNA quality. (B) In vitro transcription/translation of the p30^II ORF cloned from ACH.wt- and ACH.p30^II-immortalized cells. The p30^II ORF was amplified from both the original plasmid and immortalized-cell DNA with primers 6829 and 7550. An approximately 30-kDa product was made with the WT-derived expression vectors but was absent in the mutant (two separate clones derived from the genomic DNA were utilized).