human cdna expression library  (TaKaRa)

Journal: Arthritis Research & Therapy

Article Title: Identification of kinectin as a novel Behçet's disease autoantigen

doi: 10.1186/ar1798

Figure Lengend Snippet: cDNA clones obtained from expression library screen using a BD serum. (a) Schematic diagram of the six overlapping cDNAs aligned with published human full-length kinectin sequence (4,816 bp, GenBank accession number NM_182926 , shown on top). The open box represents the coding region of the full-length kinectin protein of ~156 kDa. cDNA inserts from the six independent clones BD41/BD44, BD481, BD42, BD47, BD482 and BD49 represent N-terminal truncations and are predicted as coiled-coil domains (hatched). ▽ represents short sequences derived from alternative mRNA splicing in the 3'-untranslated region. (b) Autoradiography of in vitro transcription and translation products of the candidate clones BD44 and BD42 labeled by [ 35 S]-methionine and analyzed on a 12.5% gel SDS-PAGE. Products of BD44 and BD42 gave major bands with the highest molecular mass of 95 kDa and 60 kDa, respectively. The lane marked as p90 represents an unrelated autoantigen used as a positive control for the in vitro translation reaction. Molecular markers are shown on the left. BD, Behçet's disease.

Article Snippet: The initial report by Hirano et al. identified kinectin by screening an aplastic anemia patient for candidate antigens using a Clontech human fetal liver cDNA expression library and it was concluded that seven out of 18 aplastic anemia patients were positive for anti-kinectin while none of the normal or disease controls had this antibody [ ].

Techniques: Clone Assay, Expressing, Sequencing, Derivative Assay, Autoradiography, In Vitro, Labeling, SDS Page, Positive Control