M13mp18 phage model as a tools of research mutagenic and cytotoxic biological and environmental compounds

*Paweł Kowalczyk¹, Karol Chalimoniuk², Agnieszka Danielak², Dorota Dziedziela², Paulina Jankowska², Marzena Kowalska², Joanna Laskowska², Marzena Rachocka², Jarosław Szczepaniak², Tomasz Walter², Paulina Strzyga², Justyna Szymańska², Mariusz Słomka², Katarzyna Zawadka², Martyna Staszewska²

Summary
Introduction. Vectors of the mp series are all derived from a recombinant M13 bacteriophage (M13mp1) that carries a short segment of E. coli DNA in its major intergenic region. This segment contains the regulatory sequences and the coding information for the first 146 amino acids of the β-galactosidase gene (lacZ). M13 is an ideal vector for obtaining single-stranded templates required for the dideoxynucleotide chain termination method of sequencing DNA and site-directed mutagenesis using synthetic oligonucleotides.
Aim. M13 phage is a usefull tools for detection of different chemical environmental compounds such as: vinyl chloride (CV), chloroacetaldehyde (CAA) and other alkylating and oxidative agents e.g. trans-4-hydroxy-2-nonenal (HNE).
Material and methods. This vector allow estimate how strong is the chemical compound; by measure the level of survival and mutation frequency in the after transfection into bacterial strains after modification of phage DNA by different compounds concentration.
Results. Modification of single stranded M13 phage DNA with HNE may inhibit in vitro DNA synthesis by T7 DNA polymerase. The most reactive base within the M13 lacZ gene was 63% of guanine residues within the examined sequence), followed by cytosine (55%), adenine (39%), and finally thymine (25%).
Conclusions. The long chain HNE adducts to DNA bases arrest DNA synthesis to all four DNA bases and cause recombination, base substitutions and frameshift mutations in ssDNA and may constitute a strong hindrance for DNA synthesis. This DNA modification may lead to mutation and finally induce tumor progression.

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