RNA POLYMERASES FROM BACTERIOPHAGE PHI 6-PHI 14 AND USE THEREOF

摘要

A polymerase protein originating from a dsRNA virus catalyzes RNA synthesis using ssRNA, dsRNA, ssDNA, or dsDNA templates. Such a polymerase can be purified from a dsRNA virus, and a protein having the amino acid sequence of sucha polymerase is useful in methods and kits for in vitro RNA synthesis. A polymerase of the invention is processive, has very high RNA-polymerization rate and does not require primer for the initiation of RNA synthesis, although it is also able to initiate RNA synthesis in the presence of a primer. Primer-independent synthesis is especially useful in amplifying RNA for quantitation of RNA species in the sample and their identification by direct sequencing. This methodology is especially useful in detecting pathogenic parasites and differences in gene expression levels associated with diseases.