Complete phage genomes can also be assembled in vitro from synthetic oligonucleotides. The entire genome of phage φX174 (5,386 bp) has been assembled in this way. Synthetic oligonucleotides are gel purified, phosphorylated, annealed, and assembled in vitro by polymerase cycle assembly (PCA). The full-length genome was amplified by PCR, digested with restriction enzymes, gel purified and circularized by ligation (phage φX174 has a closed circular genome). The assembled φX174 genome was then electroporated into E. coli and plated to examine plaques. The infectivity of synthetic phage DNA was lower than that of natural phage DNA, and this difference was attributed to PCR-generated mutations (1 per 500 bp) introduced by this method. Nonetheless, surviving phages were recovered. Due to the challenges of manipulating large DNA molecules in vitro and the potential introduction of mutations through PCR-based processes, this approach may be limited to relatively small phage genomes. However, in vitro DNA synthesis and assembly enables the introduction of arbitrary genetic changes into phage genomes more easily than recombination-based methods1.
Fig 1. Schematic diagram of the steps in the global synthesis of infectious φX174 bacteriophage from synthetic oligonucleotides. (Smith H O, et al. 2003)Creative Biolabs provide a whole-gene synthesis platform for specifying the optimal solution for phage whole-genome synthesis and assembly from synthetic oligonucleotides. We are committed to providing a proven platform and services for academic and industrial researchers focused on phage therapy. Our services are listed as follows, but are not limited to:
Competitive price: an autonomous sequencing platform with controllable cost.
High sequencing accuracy: PCR amplification errors will not accumulate, high-fidelity sequence information.
Rich experience: rich experience in whole-genome synthesis and assembly projects, especially in the leading position in phage research.
Creative Biolabs can meet the needs of customers by providing bacteriophage whole-genome synthesis and assembly from synthetic oligonucleotides services on time and on budget. We have in-depth knowledge and experience of the tools and processes involved in the phage projects. Our skilled and dedicated scientific researchers ensure that the most suitable methods and techniques are selected for your project. If necessary, please feel free to contact us.
Please kindly note that our services can only be used to support research purposes (Not for clinical use).
Creative Biolabs is a globally recognized phage company. Creative Biolabs is committed to providing researchers with the most reliable service and the most competitive price.
