Identification and sequence of the basic replication region of a broad-host-range plasmid isolated from Thiobacillus ferrooxidans

Abstract

The minimum region required for replication of the broad-host-range Thiobacillus ferrooxidans plasmid pTF-FC2 in Escherichia coli was shown to be contained on a 2.05-kilobase fragment of DNA. A 184-base-pair fragment that was required in cis for plasmid replication was identified. This region was also involved in plasmid incompatibility. Nucleotide sequencing of this region revealed three perfectly conserved 22-base-pair tandemly repeated sequences. A comparison of this region with the equivalent region of the broad-host-range plasmid R1162 showed that the repeated sequences had 60% nucleotide homology. The 106-base-pair region immediately adjacent to the repeated sequences was 75% homologous. These plasmids were compatible.