Figure 5

Optimal design of RTEm26-CTE combination element. A) Expression of pNLgag containing up-regulatory mutant RTEM26 or the wild type RTE in combination with the CTE. HeLa cells were transfected with the independent clones of indicated plasmids and analyzed for Gag expression as described in Figure 1. Standard deviations are shown. B) Organization of RTEm26-CTE element. pNLgag containing either RTEm26-CTE or the CTE-RTEm26, having the elements in reverse order separated by a 28-nt polylinker spacer, were analyzed. A spacer of 325 nt from either a synthetic HIV-1 tat gene (SP1) or from the cat gene (SP2) were inserted between RTEm26 and CTE in pNLgagRTEm26-CTE. A typical experiment is shown using the average of two to four plasmids per construct. The data are presented in % of Gag production by normalizing the values produced by pNLgagRTEm26-CTE to 100%.