Figure 5.

Demonstration of increased processivity of RTs containing K65R+L74I. Various mutant RTs were incubated with template/primer poly (rA)-oligo (dT) in the presence of 50 molar excess of trap poly (rC)-oligo (dG) and α-³²p TTP. cDNA were purified by phenol/chloroform extraction and run on a 6% polyacrylamide gel electrophoresis. Wet gels were exposed to autoradiography. cDNA fragments of different lengths and intensities are shown here. In actual autoradiograph, we were able to observe the largest cDNA bands of 72 nt, 48 nt and 54 nt in length for WT, K65R+L74V, and K65R+L74I respectively. The autoradiograph shows increased intensities of cDNA bands (13-36) synthesized with 4 and 6 μl of RT lysates of K65R+L74I viruses in comparison to K65R+L74V RT lysates (see Figure 6).