Synfacts 2020; 16(03): 0350
DOI: 10.1055/s-0039-1690344
Chemistry in Medicine and Biology
© Georg Thieme Verlag Stuttgart · New York

Total Synthesis of a Gene

Rezensent(en):
Dirk Trauner
,
Klaus-Peter Ruehmann
Khorana HG. Massachusetts Institute of Technology, Cambridge, USA
Total Synthesis of a Gene.

Science 1979;
203: 614-625
Weitere Informationen

Publikationsverlauf

Publikationsdatum:
18. Februar 2020 (online)

 

Significance

The entire gene encoding the precursor of the E. coli tyrosine tRNA was synthesized from protected mononucleotides. The final DNA fragment consists of 207 base-pairs and also includes a promotor region as well as a termination-inducing element. After successful cloning of this duplex, transcription was demonstrated in E. coli.


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Comment

First, 26 distinct oligonucleotides were synthesized using cycles of DCC-mediated coupling and basic hydrolysis to release the free 3′-OH. Remaining protecting groups of the 9- to 13-mers were cleaved and a radioactive phosphate was installed enzymatically at the 5′-OH. Ligase-mediated couplings of four to six oligonucleotides afforded six different double-stranded duplexes, which were further ligated to yield the final gene.


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