Patent ID: 6303299
Filing Date: 2001-10-16
Classification: C12N,C12Q

Abstract:
A rapid method for determining the use of different translation initiation sites of a nucleic acid encoding a protein, said method comprising:a) providing said nucleic acid, wherein the nucleic acid comprisesa first and second stop codon, said first and second stop codon being seperate and distinct, anda first and second translation initiation site, wherein said first stop codon is positioned between said first translation site and said second translation initiation site, and said second stop codon is located downstream from the second translation initiation site;b) expressing proteins encoded by the nucleic acid in the presence of a suppressor tRNA misacylated with a labelled amino acid;c) determining the total amount of protein synthesized;d) determining the total amount of labelled protein synthesized, wherein the labelled protein represents the amount of protein translated from the first translation initiation site; ande) comparing the amount of labelled protein synthesized relative to the amount of protein synthesized to determine the relative use of the first and second translation initiation sites.