Fig. 1. Hrp65-encoding clones isolated in yeast two-hybrid screening. (A) Graphical depictions of the three Hrp65 isoforms. The sequence between amino acids 1-499 is common to all isoforms, but the C-terminal sequences are variable. (B) Identification of the Hrp65 isoforms isolated in the yeast 2-hybrid screening. Plasmids isolated from each Hrp65-encoding clone were used as templates for multiplex PCR reactions with one sense oligonucleotide that is common to all known Hrp65 isoforms and two antisense oligonucleotides that discriminate between the three Hrp65 isoforms. The PCR products were separated by agarose gel electrophoresis and stained with ethidium bromide. The figures above the lanes indicate the first amino acid encoded in each cDNA. (C) Schematic representation of individual Hrp65 cDNAs isolated in the two-hybrid screening. For each clone, the first encoded amino acid downstream of the GAL4 AD is indicated to the left. The differences in length at the C-terminus indicate the isoform as determined in (B).