Table 2.

 Landmarks in the race toward the synthesis of artificial minimal genomes

References	Title	Remarks
Sekiya et al. (1979)	Total synthesis of a tyrosine suppressor tRNA gene. XVI. Enzymatic joinings to form the total 207‐base pair‐long DNA	Khorana's research group presents the last in a series of 16 papers, published along 5 years, describing the chemical synthesis of a 207‐bp tRNA gene
Nambiar et al. (1984)	Total synthesis and cloning of a gene coding for the ribonuclease S protein	The first protein‐coding gene is synthesized in the Sidney Brenner's group. Two people, working full‐time during a year and a half, were needed to synthesize this 330‐bp DNA fragment (S.A. Brenner, pers. commun.)
Stemmer et al. (1995)	Single‐step assembly of a gene and entire plasmid from large numbers of oligodeoxyribonucleotides	The first report of the synthesis of extended stretches of DNA, using a PCR‐based method for the synthesis of a 2.7‐kb plasmid from a pool of short, overlapping synthetic oligonucleotides
Cello et al. (2002)	Chemical synthesis of poliovirus cDNA: generation of infectious virus in the absence of natural template	The first chemically synthesized genome. A functional 7740‐bp poliovirus genome, with all expected biochemical and pathogenic properties was obtained
Yount et al. (2003)	Reverse genetics with a full‐length infectious cDNA of severe acute respiratory syndrome coronavirus	Viral genome assembled from cDNAs
Smith et al. (2003)	Generating a synthetic genome by whole genome assembly: phiX174 bacteriophage from synthetic oligonucleotides	Venter's group synthesizes a 5385‐bp bacteriophage genome in 2 weeks by serialized oligonucleotides assembly and amplification
Kodumal et al. (2004)	Total synthesis of long DNA sequences: synthesis of a contiguous 32‐kb polyketide synthase gene cluster	A couple of months were sufficient to synthesize a genome fragment of 32 kb, the biggest piece of man‐made DNA at that time, incorporating several genes needed for the synthesis of a pharmaceutical compound
Tian et al. (2004)	Accurate multiplex gene synthesis from programmable DNA microchips	Presents an oligonucleotide synthesis method miniaturized on photo‐programmable microfluidic chips, allowing both reduced error frequencies and lower costs. The procedure allowed the synthesis of all 21 genes that encode proteins of the E. coli 30S ribosomal subunit and to improve translation efficiency in vitro through alteration of codon bias.
Gibson et al. (2008)	Complete chemical synthesis, assembly, and cloning of a Mycoplasma genitalium genome	The complete synthesis of the 582 970‐bp genome for the pathogenic bacterium M. genitalium is reported. Venter's group used assembled chemically synthesized oligonucleotides, joined by in vitro recombination to produce intermediate assemblies, which were all cloned as bacterial artificial chromosomes in E. coli, and transferred into yeast, to assemble the full‐length genome