A Subtractive Cloning Approach to the Identification of mRNAs Specifically Expressed in Pancreatic β-cells

摘要: A polymerase chain reaction-based subtractive hybridization procedure was applied to cDNAs prepared from mouse insulinoma (βTC3) and glucagonoma (αTC2) cell lines construct a library of that are highly expressed in pancreatic β-cells. An analysis 555 randomly chosen clones the showed 80 were derived abundant mRNAs accounted for by 29 distinct sequences. Of these, 17 identical or homologous known mammalian sequence tags. Genes be β-cells represented at high frequency, namely insulin (15 clones), islet amyloid polypeptide (8 proinsulin convertase 1 (6 neuropeptide Y (2 clones). Many novel cDNA sequences relative specificity compared with other tissues, including glucagonoma, liver, kidney, brain, 3T3 fibroblasts, AtT20 corticotrophs, warrant further investigation. When combined functional immunological screening procedures, approach will useful isolation β-cell–specific molecules genetic investigations β-cell function pathology.