Multiple iterations of the M13 bacteriophage genome have been constructed by Joachim Messing and coworkers. These vectors have served as indispensable tools in the preparation, cloning, and sequencing of single-stranded DNA. Additionally, they have served as scaffolds in the construction of phage-displayed peptide libraries. Messing began by inserting the lacZα gene into the origin of replication of M13 bacteriophage, affording M13mp1. Several ensuing M13mp versions (e.g., M13mp2, M13mp6, M13mp7, etc.) were engineered to contain multiple restriction sites, creating what eventually became the polylinker of M13mp18. At the time, the sequence of each novel vector in the M13mp series was inferred from the previous vector sequence and the changes made to it; it is these sequences that are found in databases and in the literature. We suspect that some of the mutations present in M13mp18 (compared to the wild-type) arose earlier in the series that previously reported. Our goal is to determine the genomic sequences of the various M13mp vectors. Specifically, we have analyzed the whole genome sequences of M13mp1, M13mp4, and M13mp6, as well as targeted locations in M13mp5 and M13mp7. To date, we have identified a few discrepancies between the actual sequences of M13mp6 and M13mp7 compared to their reported sequences. As M13mp-based phage have a propensity to acquire compensatory mutations in phage-displayed peptide libraries, we are interested in mapping out the occurrences of earlier mutations that arose during the multi-step construction of the M13mp series. The overarching goal is to determine how various mutations may have compensated for the manipulations made during vector construction.

Genetic analysis of the M13mp bacteriophage series

2019 Spring National Meeting

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