Question

Transposon mutagenesis lab

A bacterial cell contains pRL27, but the cell does not contain the pir gene. If this bacterial cell was grown in LB broth, then describe the population of cells generated in the resulting culture and how they were generated? Be specific.

Answer 1

PIR protein is essentially required for the replication of plasmid. Therefore, if the cell lack the PIR gene pRL27 plasmid will not be able to replicate with in the cell because pRL27 plasmid is PIR protein dependent a conjugative plasmid. The pRL27 plasmid contains antibiotic resistance gene such as kanamycin (Kan) and highly active Tn5-transposase gene (TNP). Therefore, when the transformed bacteria cell pRL27 plasmid were allow to grow in a broth (selection medium with kanamycin) the bacterial population will consistently decline because in the absence of PIR gene the pRL27 plasmid will not replicate and the cell won’t be kanamycin resistance.