Development of novel genetic engineering technique in the radioresistant bacterium

Sato, Katsuya; Oba, Hirofumi*; Tu, Z.*; Narumi, Issei

To develop new shuttle vectors for species, the nucleotide sequence of the small cryptic plasmid pUE30 from was determined. The 2,467-bp plasmid possesses two open reading frames (Orf1 and Orf2). Hybrid plasmids consisting of pUE30 and pKatCAT5, which replicates in with a chloramphenicol resistance determinant, were shown to autonomously replicate in . Deletion analysis revealed that Orf2 was necessary for replication of the plasmids in . An expression plasmid that possesses the minimal promoter was constructed, and a firefly luciferase gene was successfully expressed in . The host-vector system developed in this study should prove useful in the bioremediation of radioactive waste and for the investigation of DNA repair mechanisms.