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Additional info for Houben-Weyl Methoden der organischen Chemie vol.E8d Hetarenes III, Part 4
Normally, the RNA primer is removed immediately before or after completion of replication of the circular DNA, how ever, in the presence of chlorampenicol RNA-containing, supercoiled ColEj DNA molecules accumulate. (From Blair jet. aj.. (27)). , supercoils as a result of a primer role that the RNA may be playing in the initiation of ColEj DNA synthesis. The randomness of the RNase sensitive site with respect to the two strands requires, according to this model, that either initiation of DNA synthesis is unidirectional and random with respect to the DNA strand, or synthe sis is bidirectional and initiated with short RNA fragments from both strands of the replicating mole cule.
In the case of conjugation the broken strand is transferred to the recipient cell. In the case of replication DNA poly merization would ensue, or, if already primed, per mitted to continue following the nicking event. To date, however, there is no direct evidence for a role of relaxation complexes in plasmid DNA replication or transfer and the physiological role of these com plexes remains to be determined. , replication (Kingsbury and Helinski, manuscript submitted for publication). The polAl mutant studies also showed some dependency for the DNA polymerase I enzyme for ColE 2 mainte nance.
In the experiment shown in Figure 2, the colicins were iodinated with radioactive 1 2 5 iodine prior to trypsin digestion, and the iodine con taining peptides (containing tryosine and/or histidine) were localized on fingerprint maps by autoradiography. Figure 3 shows tracings of experiments in which arginine containing peptides are specifically localized by reaction with phenanthrenequinone. 57 Prolate 11 11 Oblate lh lh Molecular Weight Axial Ratio 49 JORDAN KONISKY arginine containing peptides are shown.