Segregational Stability of the High-Copy Plasmid pUC19 in Escherichia coli Trans10 During Serial Passaging
Keywords:
pUC19, stability plasmid, Escherichia coli Trans10, high copy construction, serial passagingAbstract
The stability of plasmids is an important aspect of molecular cloning and recombinant DNA technology, directly impacting the reliability and reproducibility of plasmid-based experiments. In this work, we have studied the stability of the high-copy plasmid pUC19 in E.\ coli Trans10 during seven consecutive days of continuous serial passaging in selective (ampicillin containing) and non-selective conditions. After chemical transformation with heat shock in CaCl₂, 100 random colonies were propagated daily and the excess time of the experiment confirmed maintenance of the plasmid. All colonies were positive for pUC19 protection, regardless of antibiotic. These results suggest that the multicopy replication of pUC19 along with the genetic background of E. coli Trans10 is reliable to carry out plasmid inheritance through several generations. This work has demonstrated the stability of pUC19 as a cloning vector in Trans10 and presented an evidence that strong plasmid maintenance of it is ensured even without consistent antibiotic selection, justifying its usefulness for basic molecular biology experiments.
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