Figure 1. The MA-TC tag does not affect Gag function.

(A) Schematic diagram of HIV-1 Gag indicating the position of TC tag insertion. The amino acid sequence of the TC tag is shaded. (B) Replication kinetics of WT HIV-1 vs. the MA-TC derivative in the Jurkat T-cell line and primary MDM. Jurkat and MDM experiments were performed with pNL4-3 and pNL(AD8) molecular clones, respectively. Media were obtained every two days for RT analysis. (C) Virus release efficiency of WT vs. MA-TC. HeLa cells were transfected with WT pNL4-3 or pNL4-3/MA-TC plasmids. Transfected cells were labeled with FlAsH or DMSO (control) for 5 min 24–48 hrs posttransfection and were washed for 20 min with 300 µM EDT/PBS. The cells were then metabolically labeled with [³⁵S]Met/Cys for 2 hrs. Released virions were pelleted by ultracentrifugation, and both cell and virus lysates were immunoprecipitated with HIV-Ig and subjected to SDS-PAGE. Bands were quantified using a phosphorimager. +/− SD, n = 3.