Generation and characterization of the CA mutant library.

<p>(A) Schematic representation of the replication competent HIV-1 proviral plasmid pNHGcapNM used in this study. NotI and MluI restriction sites flank a region of Gag that includes nearly all of CA, and a few amino acids in MA. This 717 bp sequence was subjected to Genemorph II mutagenesis and subcloned into pNHGcapNM. (B) Distribution of the number of nucleotide changes in each clone for the resulting library (this includes sequences that were represented more than once). (C) Distribution of the number of amino acid changes in each clone in the library (this includes sequences that were represented more than once, nonsense mutations, mutations in MA and frameshifts). (D) Pie chart describing all of the resulting single amino acid changes. SINGLE clones were those included in further studies of the library, and the remaining clones were discarded for the following reasons: STOP (mutation resulted in a stop codon), MA (contained a mutation in the small fragment of MA included in the amplicon), FS (frameshift), DUP (duplicate mutation of clone already in library).