Kinetoplast DNA segments function as promoters in Escherichia coli cells

By means of coupled transcription-translation in Escherichia coli cell-free system, an open reading frame was found in the Crithidia oncopelti maxi-circle kDNA segment cloned in the hybrid plasmid pCo1. Subfragments from this region were tested for their ability to function as promoters in E. coli cells. For this purpose the vector pVE8 was constructed using the aminoglycoside phosphotransferase II (APTII) gene from the Tn5 transposon, and the pHC79 cosmid. After cloning of Sau3A fragments of pCo1 by insertion into pVE8 three types of plasmids containing promoter sequences were obtained. Two of these plasmids displayed promoter strength in E. coli cells greater than that of the normal promoter of the APTII gene. However the promoters found are not necessary for the coupled transcription-translation of kDNA in the E. coli cell-free system.