Cloning and Trimming of the TTR-cDNA Gene by PCR

To study the molecular basis for hereditary amyloidosis, a human adult liver cDNA was screened using PCR to isolate the normal human TTR-cDNA. Simultaneously trimming of the cDNA in both 5β- and 3β-ends was made. The PCR-product was digested with SphI and XbaI, and resulting fragments indicated successful amplification of modified TTR-cDNA, which was later cloned into vectors M13mp19 and pUC18. Positive hybridization signals were obtained in Southern blots. DNA-sequencing of the cDNA cloned in M13, revealed the successful cloning of the trimmed TTR-cDNA. A mutant TTR-cDNA with a A for G76 was observed.