Clones that were immunoreactive against the serum of patients with TIA were screened using a commercially available human aortic endothelial cell cDNA library (Uni-ZAP XR Premade Library, Stratagene, La Jolla, CA). Escherichia coli (E. coli) XL1-Blue MRF′ was infected with Uni-ZAP XR phage. Further details were described in our previously published and improved version of the immunoscreening method [18, 26, 30, 33, 34].

The monoclonalized phage cDNA clones were converted into pBluescript phagemids by in vivo excision using ExAssist helper phage (Stratagene). Plasmid DNA was obtained from the E. coli SOLR strains transformed by the phagemids. We sequenced the inserted cDNAs, followed by homologous analysis using a public database provided by the National Center for Biotechnology Information (https://blast.ncbi.nlm.nih.gov/Blast.cgi).

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