title: Discovery and characterization of a thermostable bacteriophage RNA ligase homologous to T4 RNA ligase 1.
abstract: 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 degrees 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.
authors.affiliation: Prokaria Ltd, Gylfaflot 5, 101 Reykjavik, Iceland.
authors.forename: Thorarinn
authors.initials: T
authors.lastname: Blondal
authors.forename: Sigridur H
authors.initials: SH
authors.lastname: Hjorleifsdottir
authors.forename: Olafur F
authors.initials: OF
authors.lastname: Fridjonsson
authors.forename: Arnthor
authors.initials: A
authors.lastname: Aevarsson
authors.forename: Sigurlaug
authors.initials: S
authors.lastname: Skirnisdottir
authors.forename: Anna Gudny
authors.initials: AG
authors.lastname: Hermannsdottir
authors.forename: Gudmundur O
authors.initials: GO
authors.lastname: Hreggvidsson
authors.forename: Albert Vernon
authors.initials: AV
authors.lastname: Smith
authors.forename: Jakob K
authors.initials: JK
authors.lastname: Kristjansson
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000201:enzymology* / Q000235:genetics; D001709:Biotechnology; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded / Q000378:metabolism; D004795:Enzyme Stability; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D012313:RNA / Q000378:metabolism; D011130:RNA Ligase (ATP) / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D045245:Rhodothermus / Q000821:virology*; D016415:Sequence Alignment; D013379:Substrate Specificity; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D004277:DNA, Single-Stranded; D014764:Viral Proteins; D012313:RNA; D000255:Adenosine Triphosphate; D011130:RNA Ligase (ATP); C477285:bacteriophage T4 RNA ligase 1