METHODS FOR PREPARING NUCLEOTIDE INTEGRASES

摘要

The present invention provides new, improved and easily manipulable methods for making nucleotide integrases. The present method also relates to a nucleotide integrase and an improved method for making RNA-protein complexes for use in preparing nucleotide integrases in vitro. The nucleotide integrase is prepared by introducing a DNA molecule which comprises a group II intron DNA sequence into a host cell. The group II intron DNA sequence is then expressed in the host cell such that RNP particles having nucleotide integrase activity are formed in the cell. Such RNP particles comprise an excised group II intron RNA encoded by the introduced DNA molecule and a group II intron-encoded protein encoded by the introduced DNA molecule. Thereafter the nucleotide integrase is isolated from the cell. In another embodiment, the nucleotide integrase is prepared by combining in vitro an excised group II intron RNA, hereafter 'exogenous RNA', with group II intron-encoded protein. In another embodiment, the nucleotide integrase is prepared by combining 'exogenous RNA' with an RNA-protein complex which comprises a group II intron-encoded protein.