Chemical ligation of DNA: the first non-enzymatic assembly of a biologically active gene. |
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| Authors: | Z A Shabarova I N Merenkova T S Oretskaya N I Sokolova E A Skripkin E V Alexeyeva A G Balakin A A Bogdanov |
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| Affiliation: | Belozersky Laboratory of Molecular Biology and Bioorganic Chemistry, Moscow State University, USSR. |
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| 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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