The M13mp vectors have served as indispensable tools in the preparation, cloning, and sequencing of single-stranded DNA. Joachim Messing and coworkers constructed M13mp1 by inserting the lacZα gene into the origin of replication of M13 bacteriophage. Several ensuing M13mp versions contained iterations of what eventually became the polylinker of M13mp18. Based on sequences gathered from online databases, it appears that during the engineering of the polylinker, it was understood that the rest of the phage genome generally remained unaltered. However, we have recently sequenced wild-type M13 and a few of the M13mp series and found several mutations scattered throughout the genome. These mutations may have spontaneously arisen in order to compensate in part for the replication defect caused by lacZα insert in the ori, which is known decrease the propagation rate of the phage. Our goal is to map the mutations in the intermediate vectors from M13mp1 to M13mp18 in order to pinpoint at which step each one arose. Based on our findings to date, the majority of the mutations appear to have emerged between M13mp6 and M13mp18, and further sequencing promises to elucidate the genetic progression of the M13mp vectors. Additionally, we have demonstrated that various M13mp phage have different propagation rates, falling between those of wild-type M13 and M13mp18. Our goal is to link each mutation to its effect on the propagation rate of the phage in order to infer whether it actually compensates for the lacZα insert, or rather is inconsequential or detrimental to the phage.

Compensatory mutations in the M13mp phage series

255th National Meeting (2018)

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