Screening of augmenter of liver regeneration-binding proteins by yeast-two hybrid technique

OBJECTIVE: To investigate the biological function of augmenter of liver regeneration (ALR), we usedyeast-two hybrid technique to detect proteins in hepatocytes interacting with ALR.METHODS: ALR bait plasmid was constructed by using yeast-two hybrid system 3, then transformedinto yeast AH109. The transformed yeast was mated with yeast Y187 containing liver cDNA libraryplasmid in a 2×YPDA medium. Diploid yeast was plated on a synthetic dropout nutrient medium(SD/-Trp-Leu-His-Ade) containing x-α-gal for selection and screening. After extracting and sequencingof the plasmid from blue colonies. Analysis was performed by bioinformatics.RESULTS: Of 36 colonies sequenced, 14 are metallothionein, 12 albumin, and 3 selenoprotein P. Onecolony is a new gene with unknown function.CONCLUSION: The successful cloning of gene of ALR interacting protein has paved the way forstudying the physiological function of ALR and associated proteins.