Gene synthesis by circular assembly amplification.
Here we report the development of a gene-synthesis technology, circular assembly amplification. In this approach, we first constructed exonuclease-resistant circular DNA via simultaneous ligation of oligonucleotides. Exonuclease- and subsequent mismatch cleaving endonuclease-mediated degradation of the resulting ligation mixture eliminated error-rich products, thereby substantially improving gene-synthesis quality. We used this method to construct genes encoding a small thermostable DNA polymerase, a highly repetitive DNA sequence and large (>4 kb) constructs.
Bang
Duhee
D
Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA. dbang@genetics.med.harvard.edu
Church
George M
GM
D004270:DNA, Circular / Q000235:genetics*; D005796:Genes / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics*
D004270:DNA, Circular; D011994:Recombinant Proteins