cDNA Cloning of Inducer Reactive Protein in Adult T-Cell Leukemia Cells by Applying the Fuzzy Set Cloning Theory for Sequencing Defuzzification

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Fuzzy set cloning (FSC) was performed to detect cDNAs that react to gene expression regulator molecules such ascorbic acid (VC) and hydrocortisone. Our target was proteoglycan involved in tissue construction as a component of extracellular matrices. While proteoglycan is not usually produced in hematopoietic cells, MT1 and MT2 cell-derived cell lines of adult T-cell leukemia cells (ATL cells) expressed 100 k dalton (kDa) proteglycan. We performed FSC using a partially purified anti-100 kDa protein antibody as the fuzzy set antibody (FSAb), sequenced the obtained clones, and defuzzified them as new clones using computer queries of the GenBank and EMBL databases. A second defuzzification was performed by analyzing mRNA levels by reverse transcription (RT)-PCR analysis between peripheral lymphocytes and MT1 cells after induction of gene expression by ascorbic acid and hydrocortisone. We were able to obtain functional new cDNAs that exist specifically in ATL cells but not in normal peripheral lymphocytes. Moreover, we were able to define the promoter functions as inducible by VC, hydrocortisone or as none-responsive genes.
バイオメディカル・ファジィ・システム学会の論文