title: Whole-genome synthesis and characterization of viable S13-like bacteriophages.
abstract: BACKGROUND
  Unprecedented progresses in high-throughput DNA sequencing and de novo gene synthesis technologies have allowed us to create living organisms in the absence of natural template.
  
  
  RESULTS
  The sequence of wild-type S13 phage genome was downloaded from GenBank. Two synonymous mutations were introduced into wt-S13 genome to generate m1-S13 genome. Another mutant, m2-S13 genome, was obtained by engineering two nonsynonymous mutations in the capsid protein coding region of wt-S13 genome. A chimeric phage genome was designed by replacing the F capsid protein open reading frame (ORF) from phage S13 with the F capsid protein ORF from phage G4. The whole genomes of all four phages were assembled from a series of chemically synthesized short overlapping oligonucleotides. The linear synthesized genomes were circularized and electroporated into E.coli C, the standard laboratory host of S13 phage. All four phages were recovered and plaques were visualized. The results of sequencing showed the accuracy of these synthetic genomes. The synthetic phages were capable of lysing their bacterial host and tolerating general environmental conditions. While no phenotypic differences among the variant strains were observed when grown in LB medium with CaCl(2), the S13/G4 chimera was found to be much more sensitive to the absence of calcium and to have a lower adsorption rate under calcium free condition.
  
  
  CONCLUSIONS
  The bacteriophage S13 and its variants can be chemically synthesized. The major capsid gene of phage G4 is functional in the phage S13 life cycle. These results support an evolutional hypothesis which has been proposed that a homologous recombination event involving gene F of quite divergent ancestral lineages should be included in the history of the microvirid family.
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: Yuchen
authors: Y
authors: Liu
authors: Yonghua
authors: Y
authors: Han
authors: Weiren
authors: W
authors: Huang
authors: Yonggang
authors: Y
authors: Duan
authors: Lisha
authors: L
authors: Mou
authors: Zhimao
authors: Z
authors: Jiang
authors: Pingping
authors: P
authors: Fa
authors: Jun
authors: J
authors: Xie
authors: Ruiying
authors: R
authors: Diao
authors: Yuanbin
authors: Y
authors: Chen
authors: Yiwang
authors: Y
authors: Ye
authors: Ruilin
authors: R
authors: Yang
authors: Jing
authors: J
authors: Chen
authors: Xiaojuan
authors: X
authors: Sun
authors: Zesong
authors: Z
authors: Li
authors: Aifa
authors: A
authors: Tang
authors: Yaoting
authors: Y
authors: Gui
authors: Zhiming
authors: Z
authors: Cai
mesh_terms: D000327:Adsorption; D001435:Bacteriophages / Q000235:genetics*; D002118:Calcium / Q000737:chemistry; D018274:Electroporation; D004926:Escherichia coli / Q000378:metabolism / Q000821:virology; D022761:Gene Transfer, Horizontal; D005818:Genetic Engineering / Q000379:methods; D005819:Genetic Markers; D016680:Genome, Bacterial; D016679:Genome, Viral; D006863:Hydrogen-Ion Concentration; D008957:Models, Genetic; D009154:Mutation; D016366:Open Reading Frames; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D013696:Temperature; D014466:Ultraviolet Rays
chemical_list: D005819:Genetic Markers; D002118:Calcium