in a konica srx 101a medical film processor taiwan the developed x ray film was scanned by arcus 1200 agfa scanner taiwan and scanned images were analyzed using genepix pro 4 0 axon instruments union city calif signal intensities were defined as median pixel intensity following subtraction of local median background signal the peptide array was stripped with restore u2122 western blot stripping buffer pierce rockford ill according to the procedure recommended by the manufacturer before it was probed with a different anti plpe antibody this was repeated several times with anti plpe antibodies obtained from different sources or purified in varieties of ways the determination of the epitope map of pipe was attempted in two steps the first approach involved the localization of the general area of immunogenic or immunodominant epitopes by deleting specific regions of plpe from both the n terminus and c terminus by pcr with the help of specific primers a total of 6 plasmid constructs carrying the plpe gene with varying degrees of deletions were made the cloning of three of these constructs that carry deletions from the n termini of plpe psac63 psac64 and psac65 is described above three additional plasmids viz psac30 psac31 and psac32 that carry 106 rplpe u03b4c106 96 rplpe u03b4c96 and 86 rplpe u03b4c86 amino acid deletions on the c terminus of plpe respectively were designed and constructed the reverse primers used to introduce these deletions on the 3 u2032 end of the plpe gene in the latter constructs were hnplp 1 5 u2032 gataagcttttaccgtgcggcaaattc 3 u2032 seq id no 7 hnplp 2 5 u2032 aaaaagcttttatttaatttctacatc 3 u2032 seq id no 8 and hnplp 3 5.