Abstract: Stable, constitutively expressed, chromosomal fluorescent transcriptional fusions in bacterial pathogens and methods of using the same to screen candidate compounds for anti-bacterial efficacy.

Abstract: The present invention includes live strain of Francisella tularensis wherein a gene selected from the group consisting of priA and purA is inactivated. The present invention further includes a pharmaceutical composition comprising one or more live strains of Francisella tularensis according to the present invention and a pharmaceutically acceptable carrier. The present invention further includes a method of using one or more live strains of Francisella tularensis according to the present invention to confer immunity against a virulent strain of Francisella tularensis. The method comprises administering an effective amount of one or more live strains of Francisella tularensis according to the present invention or a pharmaceutical composition comprising one or more live Francisella tularensis strains to an animal such that an immune response is produced in the animal.

Abstract: The present invention includes live strain of Francisella tularensis wherein a gene selected from the group consisting of priA and purA is inactivated. The present invention further includes a pharmaceutical composition comprising one or more live strains of Francisella tularensis according to the present invention and a pharmaceutically acceptable carrier. The present invention further includes a method of using one or more live strains of Francisella tularensis according to the present invention to confer immunity against a virulent strain of Francisella tularensis. The method comprises administering an effective amount of one or more live strains of Francisella tularensis according to the present invention or a pharmaceutical composition comprising one or more live Francisella tularensis strains to an animal such that an immune response is produced in the animal.

Abstract: Stable, constitutively expressed, chromosomal fluorescent transcriptional fusions in bacterial pathogens and methods of using the same to screen candidate compounds for anti-bacterial efficacy.

Abstract: Provided herein are nucleic acid constructs and methods for producing or enhancing the production of group II intron RNP particles in eukaryotic cells. The present methods comprise introducing at least one nucleic acid construct comprising a nucleic acid encoding a modified or wild type group II intron RNA and a wild-type or modified group II intron-encoded protein into the eukaryotic cell, and maintaining the cell under conditions that allow for expression of the group II intron RNA and the group II intron-encoded protein in the cell. The nucleic acid encoding the group II intron RNA is operably linked to an RNA polymerase I, an RNA polymerase II, or an RNA polymerase III promoter, and the nucleic acid encoding the group II intron-encoded protein is operably linked to an RNA polymerase II promoter. In certain embodiments, a subcellular localization signal is attached to the group II intron-encoded protein.

Abstract: Methods for preparing nucleotide integrases are provided. The nucleotide integrases are prepared by combining in vitro an excised, group II intron RNA, referred to hereinafter as "exogenous RNA", with a group II intron-encoded protein. The exogenous RNA is prepared by in vitro transcription of a DNA molecule which comprises a group II intron sequence. In one embodiment, the group II intron-encoded protein is made by introducing into a host cell a DNA molecule that comprises at least the open reading frame sequence of a group II intron and then expressing the open reading frame sequence in the host cell. The DNA molecule may comprise the open reading frame sequence operably linked to a promoter, preferably an inducible promoter. Thereafter, the cell is fractionated and the protein is recovered and combined in vitro with the exogenous RNA to provide RNP particles having nucleotide integrase activity.

Abstract: The present invention provides new, improved, and easily manipulable methods for making nucleotide integrases. In one embodiment, 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 exiced 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, referred to hereinafter as "exogenous RNA", with a group II intron-encoded protein.