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Figure 1 | Retrovirology

Figure 1

From: In vivo analysis of Nef’s role in HIV-1 replication, systemic T cell activation and CD4+ T cell loss

Figure 1

HIV-1JRCSF with a truncated nef. a Upper panel schematic representation of wild type JRCSF nef (WT JRCSF) is presented. Nucleotides 8784–9434 in NCBI accession number, M38429, represent the nef coding sequence. PPT polypurine tract. Lower panel, a schematic presentation of nef with two deletions (JRCSFNefdd). A 114 bp deletion 5′ of the PPT from 8919 to 9032 inclusive was introduced into nef. The downstream sequence was shifted to the +1 reading frame by cutting with XhoI and filling in with Klenow (“Methods”). The 293 bp deletion 3′ of the PPT shifted downstream nef sequence to reading frame to +2. b The proviral clones for JRCSF and JRCSFNefdd were transfected into 293T cells and after 2 days Nef and Env expressions assessed by Western blots, GADPH is a loading control. c Replication competence of JRCSFNefdd was not diminished by loss of Nef as determined by p24gag production in CEM cells expressing CCR5. Cells were infected at 1 × 105 TCIU at an MOI 0.01 and the production of p24gag was followed for 21 days.

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