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Expression of Nef from recombinant HIV-1. The sequence-verified nef alleles from LTNPs, AIDS patients, and ACs were inserted into the XhoI/NotI site of pNL-Not. The infectious DNAs were transfected into 293T cells to propagate viruses. The cell lysates were analyzed by being immunoblotted with a pool of anti-Nef monoclonal antibodies.
