Structural Biochemistry/T4 RNA Ligase
T4 RNA Ligase catalyzes the ligation of a 5' phosphoryl-terminated nucleic acid donor to a 3' hydroxyl-terminated nucleic acid acceptor through the formation of a 3'→5' phosphodiester bond, with hydrolysis of ATP to AMP and PPi. Substrates include single-stranded RNA and DNA as well as dinucleoside pyrophosphates. This ligase can be formed from Purified E. coli strain ER2497 which contains the plasmid pRF-E35. This enzyme is used for the labeling the 3’-terminus of RNA with a 5’-[32P]. It functions as the inter- and intramolecular joining of the RNA and DNA molecules. It can also be used to synthesize single stranded oligodeoxyribo-nucleotides as well as incorporating additional amino acids into proteins.