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Nucleic Acids Res ; 45(7): 3997-4005, 2017 04 20.
Artigo em Inglês | MEDLINE | ID: mdl-28158820

RESUMO

Biological evolution resulted in a homochiral world in which nucleic acids consist exclusively of d-nucleotides and proteins made by ribosomal translation of l-amino acids. From the perspective of synthetic biology, however, particularly anabolic enzymes that could build the mirror-image counterparts of biological macromolecules such as l-DNA or l-RNA are lacking. Based on a convergent synthesis strategy, we have chemically produced and characterized a thermostable mirror-image polymerase that efficiently replicates and amplifies mirror-image (l)-DNA. This artificial enzyme, dubbed d-Dpo4-3C, is a mutant of Sulfolobus solfataricus DNA polymerase IV consisting of 352 d-amino acids. d-Dpo4-3C was reliably deployed in classical polymerase chain reactions (PCR) and it was used to assemble a first mirror-image gene coding for the protein Sso7d. We believe that this d-polymerase provides a valuable tool to further investigate the mysteries of biological (homo)chirality and to pave the way for potential novel life forms running on a mirror-image genome.


Assuntos
DNA Polimerase beta/genética , DNA/biossíntese , Proteínas Arqueais/genética , DNA/química , DNA Polimerase beta/síntese química , DNA Polimerase beta/metabolismo , Proteínas de Ligação a DNA/genética , Estabilidade Enzimática , Mutação , Estereoisomerismo , Sulfolobus solfataricus/enzimologia , Temperatura
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