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. 2001 Feb;75(4):1834–1841. doi: 10.1128/JVI.75.4.1834-1841.2001

FIG. 4.

FIG. 4

Effect of altered Gag/Gag-Pol ratios on virion RNA dimerization. The impact of various Gag/Gag-Pol ratios on genomic RNA dimerization was determined using melting curve and electrophoretic analysis of wild-type (WT) and mutant dimers. Virion RNA was resuspended in RNA dimerization buffer and heat denatured for 10 min at the indicated temperatures. Dimers and monomers were electrophoresed in a 1% native agarose gel and probed with an HIV-1 riboprobe as described in Materials and Methods. RNA dimerization analysis involved wild-type HIV-1 and mutant viruses derived from cells cotransfected with wild-type and GP PR(+) expression vectors to achieve Gag/Gag-Pol ratios of 20:1 to 20:21 (A), cells cotransfected with wild-type and GP PR(−) expression vectors to achieve Gag/Gag-Pol ratios of 20:3.5 to 20:21 (B), and cells transfected with wild-type proviral DNA, transfected with a PR-defective full-length HIV-1 Pr-defective plasmid [PR(−)] for the production of PR-defective immature HIV-1 particles, and cotransfected with wild-type GP PR(−) to achieve a Gag/Gag-Pol ratio of 20:21 (C). U, unheated.