title: Chemical synthesis of bacteriophage G4.
abstract: BACKGROUND
  Due 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.
  
  
  RESULTS
  Full-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.
  
  
  CONCLUSIONS
  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.
authors: Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics, Institute of Urology, Peking University Shenzhen Hospital, Shenzhen PKU-HKUST Medical Center, Shenzhen, China.
authors: Ruilin
authors: R
authors: Yang
authors: Yonghua
authors: Y
authors: Han
authors: Yiwang
authors: Y
authors: Ye
authors: Yuchen
authors: Y
authors: Liu
authors: Zhimao
authors: Z
authors: Jiang
authors: Yaoting
authors: Y
authors: Gui
authors: Zhiming
authors: Z
authors: Cai
mesh_terms: D004262:DNA Restriction Enzymes / Q000378:metabolism; D004926:Escherichia coli / Q000821:virology; D016679:Genome, Viral / Q000235:genetics; D008854:Microscopy, Electron; D017915:Microvirus / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure; D016133:Polymerase Chain Reaction; D058615:Synthetic Biology / Q000379:methods*
chemical_list: D004262:DNA Restriction Enzymes