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. 2018 Dec 19;13(12):e0208345. doi: 10.1371/journal.pone.0208345

Fig 1. Rapid Immunogen Selection (R.I.S.) method.

Fig 1

(A) Random mutations were introduced into HIV-1 env AC10_29 sequence by mutagenic PCR. (B) A library of chimeric HIV virions was generated by cloning randomly mutated envelopes into pNL4-3 using XbaI and NotI restriction sites and transient transfection into 293T cells. (C) Virion library was used in an enhanced virion capture assay modified from Leaman et al as indicated in the Materials and Methods section (41). (D) RNA from captured virions was extracted, retrotranscribed into DNA and amplified by nested RT-PCR. (E and B) A new library of chimeric virions was generated by transient transfection of the env amplified product out of captured virions and cloned into NL4-3 proviral DNA.