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The manufacturing of viruses in the test tube begins with the observations that viral capsid proteins undergo self-assembly when ionic and pH conditions are suitable. Components of Escherichia coli phage, such as T4 and λ, assemble spontaneously to form whole phage particles, and components of icosahedral viruses in the presence or absence of nucleic acid form spherical structures. Viral RNA was copied by the reverse transcriptase into cDNA and expressed on a plasmid with the formation of intact virus. By 2002, poliovirus was synthesized from full-length cDNA. With the progress in technology for the synthesis of long stretches of DNA and improvements in sequencing, it was a natural next step to construct a virus in a cell-free system using commercially available DNA. Poliovirus and Qβ bacteriophage were synthesized in vitro; similar techniques were used to sequence and reconstitute the 1918 flu pandemic virus, and to modify the host range of avian influenza virus. These latter experiments were criticized as being dangerous to humans and to the environment, and new regulations were introduced to control such experiments.
KeywordsInfluenza Virus H5N1 Virus Avian Influenza Virus Yersinia Pestis Francisella Tularensis
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