RNA-Dependent RNA Polymerase, Methods And Kits For The Amplification And/Or Labelling Of RNA

A method for the amplification of RNA using an RNA-dependent RNA polymerase having a right hand conformation and an amino acid sequence comprising the following sequence motifs:

• XXDYS
• GXPSG
• YGDD
• XXYGL
• XXXXFLXRXX
• wherein the amino acids in the sequence motifs are identified as:
• D:
• aspartate
• Y:
• tyrosine
• S:
• serine
• G:
• glycine
• P:
• proline
• L:
• leucine
• F:
• phenylalanine
• R:
• arginine
• X:
• any amino acid
• comprising the steps of:
• a. annealing of the RNA-dependent RNA polymerase to the RNA template in the presence or absence of an oligonucleotide primer;
• b. transcribing the RNA template into antisense-RNA by the RNA-dependent RNA polymerase;
• c. separating the RNA/antisense-RNA duplex into individual strands.

A method for the labelling of RNA using RNA-dependent RNA polymerase having a right hand conformation and an amino acid sequence comprising the following sequence motifs:

• XXDYS
• GXPSG
• YGDD
• XXYGL
• XXXXFLXRXX
• wherein the amino acids in the sequence motifs are identified as:
• D:
• aspartate
• Y:
• tyrosine
• S:
• serine
• G:
• glycine
• P:
• proline
• L:
• leucine
• F:
• phenylalanine
• R:
• arginine
• X:
• any amino acid;
• comprising the steps of:
• a. annealing the RNA-dependent RNA polymerase to the RNA to be labelled.
• b. adding at least one nucleotide to the 3′-end of the RNA to be labelled.

A kit for the amplification of RNA comprising:

• an RNA-dependent RNA polymerase having a right hand conformation and an amino acid sequence comprising the following sequence motifs:
• XXDYS
• GXPSG
• YGDD
• XXYGL
• XXXXFLXRXX
• wherein the amino acids in the sequence motifs are identified as:
• D:
• aspartate
• Y:
• tyrosine
• S:
• serine
• G:
• glycine
• P:
• proline
• L:
• leucine
• F:
• phenylalanine
• R:
• arginine
• X:
• any amino acid;
• a reaction buffer;
• NTPs;
• optionally, RNase inhibitor;
• optionally, stop solution.

A kit for labelling RNA comprising:

• an RNA-dependent RNA polymerase having a right hand conformation and an amino acid sequence comprising the following sequence motifs:
• XXDYS
• GXPSG
• YGDD
• XXYGL
• XXXXFLXRXX
• wherein the amino acids in the sequence motifs are identified as:
• D:
• aspartate
• Y:
• tyrosine
• S:
• serine
• G:
• glycine
• P:
• proline
• L:
• leucine
• F:
• phenylalanine
• R:
• arginine
• X:
• any amino acid;
• a reaction buffer;
• NTPs;
• optionally, RNase inhibitor;
• optionally, stop solution.