Nucleotide sequence of an incompatibility region of mini-Rts1 that contains five direct repeats.
The plasmid mini-Rts1, consisting of an EcoRI/HindIII fragment of about 1.8 kilobases (kb), contains an incompatibility determinant in its EcoRI/AccI region (0.5 kb). The nucleotide sequence of this incompatibility fragment was determined. A most striking feature of the sequence is the presence of five 24-base pair direct repeats. Four out of the five repeating units, which are contained in a 0.2-kb EcoRI/HincII fragment, were cloned en bloc in pACYC184 and found to express Rts1-specific incompatibility. In addition, the copy number of the mini-Rts1 plasmid appeared to be increased threefold upon removal of the 0.2-kb incompatibility region (incI) from the plasmid. This deletion derivative of mini-Rts1, as well as mini-Rts1, was maintained stably at 37 degrees C, but was cured at a high frequency at 42 degrees C. A possible role of the repeated nucleotide sequence was discussed. By subcloning the mini-Rts1 DNA, a second inc determinant (incII) was found on the AccI fragment, which is contiguous to the 0.5-kb EcoRI/AccI fragment.
D001483:Base Sequence; D004261:DNA Replication; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial; D015246:Deoxyribonuclease EcoRI; D004926:Escherichia coli / Q000235:genetics; D010957:Plasmids; D012091:Repetitive Sequences, Nucleic Acid; D013696:Temperature
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes; D015246:Deoxyribonuclease EcoRI