A Novel Approach for Obtaining acDNA Clone that Codes Functional Protein by Applying the Fuzzy Set Cloning Theory

概要

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The Fuzzy set cloning (FSC) theory was developed to identify new functional protein(s) easily and rapidly. Characteristically, in FSC the functional target protein of interest is reported as a cDNA sequence. Using FSC, most of the proteins, if they configure mass to function, are "harvested" in a "net" using fuzzy set antibody (FSAb). The cDNAs are then subjected to a specific selection procedure called defuzzification (DeFuzzy). Here, the most useful fast procedure with explicit object function should be selected for DeFuzzy. The defuzzified clones are sequenced and BLAST / IDEAS program is used to determine whether they represent a new or useful functional protein by comparing the sequences to those in the EMBL and GenBank database. We focused on inducer proteins of rat liver fibrosis after injecting the animals with carbon tetrachloride (CCl_4). Using antibody against basement membrane as FSAb, 32 positive λgt11 cDNA clones were obtained. For DeFuzzy, gene expression levels were analyzed on timed Northern blots. Sequencing of a clone coding mRNA expressed in the early phase after CCl_4 treatment identified human profilin. The profilin peptide antibody stained fibrogenic areas in cirrhotic livers from rats.
バイオメディカル・ファジィ・システム学会の論文
1996-08-08