Discovery and characterization of a thermostable bacteriophage RNA ligase homologous to T4 RNA ligase 1.
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.
Blondal
Thorarinn
T
Prokaria Ltd, Gylfaflot 5, 101 Reykjavik, Iceland.
Hjorleifsdottir
Sigridur H
SH
Fridjonsson
Olafur F
OF
Aevarsson
Arnthor
A
Skirnisdottir
Sigurlaug
S
Hermannsdottir
Anna Gudny
AG
Hreggvidsson
Gudmundur O
GO
Smith
Albert Vernon
AV
Kristjansson
Jakob K
JK
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*
D004277:DNA, Single-Stranded; D014764:Viral Proteins; D012313:RNA; D000255:Adenosine Triphosphate; D011130:RNA Ligase (ATP); C477285:bacteriophage T4 RNA ligase 1