Abstract
BackgroundDue to recent leaps forward in DNA synthesis and sequencing technology, DNA manipulation has been extended to the level of whole-genome synthesis. Bacteriophages occupy a special niche in the micro-organic ecosystem and have potential as a tool for therapeutic agent. The purpose of this study was to carry out chemical synthesis of the bacteriophage G4 and the study of its infectivity.Methodology/Principal FindingsFull-sized genomes of bacteriophage G4 molecules were completed from short overlapping synthetic oligonucleotides by direct assembly polymerase chain reaction and ligase chain reaction followed by fusion polymerase chain reaction with flanking primers. Three novel restriction endonuclease sites were introduced to distinguish the synthetic G4 from the wild type. G4 particles were recovered after electroporation into Escherichia coli and were efficient enough to infect another strain. The phage was validated by electron microscope. Specific polymerase chain reaction assay and restriction analyses of the plaques verified the accuracy of the chemical synthetic genomes.ConclusionsOur results showed that the bacteriophage G4 obtained is synthetic rather than a wild type. Our study demonstrated that a phage can be synthesized and manipulated genetically according to the sequences, and can be efficient enough to infect the Escherichia coli, showing the potential use of synthetic biology in medical application.
Highlights
Bacteriophages, being viruses that infect bacteria, have played an important role in underpinning the development and advancement of the biosciences since the dawn of molecular biology
Our results showed that the bacteriophage G4 obtained is synthetic rather than a wild type
Our study demonstrated that a phage can be synthesized and manipulated genetically according to the sequences, and can be efficient enough to infect the Escherichia coli, showing the potential use of synthetic biology in medical application
Summary
Bacteriophages, being viruses that infect bacteria, have played an important role in underpinning the development and advancement of the biosciences since the dawn of molecular biology. Bacteriophages, first discovered around 1915, occupy a special place in viral biology. They are perhaps the best understood viruses. Their genome being less than 10,000 bases long is amenable to genetic alterations at the level of whole genome synthesis thanks to the recent advance made in DNA synthesis and sequencing technology. Due to the advances in synthetic biology, it is possible to obtain large segments of synthetic DNA, assemble them into entire genomes of infectious agents, and boot them to life. Cello J et al [1] achieved the first chemical synthesis of a DNA (7,500 bp) corresponding to the entire genome of poliovirus. The purpose of this study was to carry out chemical synthesis of the bacteriophage G4 and the study of its infectivity
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
