Genetically Engineered Microorganisms and Their Impact on Human Health

Figure 1

Workflow of phage genome engineering strategies. “Genome assembly and rebooting” is kind of synthetic method and uses phage genome that assembles by smaller and then does the overlapping of DNA fragments through two ways, Gibson method and transformation associated recombination (TAR), using yeast. DNA synthetic transformed into the Gram-negative bacterium (e. coli) or Gram-positive bacterium (L form) to rebooting phage genome. Finally, phages replicate and infect the host and then followed by lysis which, for Gram-negative bacteria, perform by chloroform lysis while it has not effect on Gram-positive bacteria. Therefore, L form bacteria is using which hypotonic pressure select for cell lysis occurred and mutated (or engineered phages) release. Recombination-based methods are divided into two in vivo recombination and BRED and Crisper-Cas based method. Edited templates which were infected in host cell recombine through phage genome replication, and then, mixture of wild-type and recombinant phages was produced. In contrast to synthetic biology method, in last step of BRED and in vivo recombination method, screening is conducted to select certain and appropriate strain by plaque screening method.