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. 1991 Aug 11;19(15):4247-51.
doi: 10.1093/nar/19.15.4247.

Chemical ligation of DNA: the first non-enzymatic assembly of a biologically active gene

Z A Shabarova  1 , I N Merenkova, T S Oretskaya, N I Sokolova, E A Skripkin, E V Alexeyeva, A G Balakin, A A Bogdanov
Affiliations
Free PMC article

Chemical ligation of DNA: the first non-enzymatic assembly of a biologically active gene

Z A Shabarova et al. Nucleic Acids Res. .
Free PMC article

Abstract

An artificial gene comprising 183 base pairs has been assembled by template-directed condensation of 35- to 53-membered oligodeoxyribo nucleotides with cyanogen bromide as a condensing agent. The reaction is complete within several minutes at 0 degrees C in buffer. The resulting mini-gene was cloned and expressed in vivo and in vitro. We have also found that the polymerase inhibition technique (toe-printing) is a good way to ascertain that translation initiation complexes form in the case of single-stranded DNAs as well. Thus, along with the fully chemical assembly of synthetic genes, a rapid and sufficiently reliable method for determining their ribosome-binding properties was developed.

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References

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