Discovery and characterization of a thermostable bacteriophage RNA ligase homologous to T4 RNA ligase 1

• Published in: Nucleic Acids Research
• Main Authors: Blondal, Thorarinn, Hjorleifsdottir, Sigridur H., Fridjonsson, Olafur F., Ævarsson, Arnthor, Skirnisdottir, Sigurlaug, Hermannsdottir, Anna Gudny, Hreggvidsson, Gudmundur O., Smith, Albert Vernon, Kristjansson, Jakob K.
• Format: Text
• Language: English
• Published: Oxford University Press 2003
• Subjects: Articles; Iceland
• Online Access: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC291858; http://www.ncbi.nlm.nih.gov/pubmed/14654700; https://doi.org/10.1093/nar/gkg914

Thermophilic viruses represent a novel source of genetic material and enzymes with great potential for use in biotechnology. We have isolated a number of thermophilic viruses from geothermal areas in Iceland, and by combining high throughput genome sequencing and state of the art bioinformatics we have identified a number of genes with potential use in biotechnology. We have also demonstrated the existence of thermostable counterparts of previously known bacteriophage enzymes. Here we describe a thermostable RNA ligase 1 from the thermophilic bacteriophage RM378 that infects the thermophilic eubacterium Rhodothermus marinus. The RM378 RNA ligase 1 has a temperature optimum of 60–64°C and it ligates both RNA and single-stranded DNA. Its thermostability and ability to work under conditions of high temperature where nucleic acid secondary structures are removed makes it an ideal enzyme for RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE), and other RNA and DNA ligation applications.