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Nucleic Acids Res ; 31(24): 7247-54, 2003 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-14654700

RESUMO

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.


Assuntos
Bacteriófagos/enzimologia , RNA Ligase (ATP)/metabolismo , Rhodothermus/virologia , Proteínas Virais/metabolismo , Trifosfato de Adenosina/metabolismo , Sequência de Aminoácidos , Bacteriófagos/genética , Biotecnologia , Clonagem Molecular , DNA de Cadeia Simples/metabolismo , Estabilidade Enzimática , Temperatura Alta , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , RNA/metabolismo , RNA Ligase (ATP)/química , RNA Ligase (ATP)/genética , RNA Ligase (ATP)/isolamento & purificação , Alinhamento de Sequência , Especificidade por Substrato
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