Surface-initiated enzymatic polymerization of DNA.
We describe a technique to synthesize DNA homopolymers on a surface using surface-initiated enzymatic polymerization (SIEP) with terminal deoxynucleotidyl transferase (TdTase), an enzyme that repetitively adds mononucleotides to the 3'-end of oligonucleotides. The thickness of the synthesized DNA layer was found to depend on the deoxymononucleotide monomer, in the order of dATP > dTTP > dGTP approximately dCTP. In addition, the composition and the surface density of oligonucleotide initiators were also important in controlling the extent of DNA polymerization. The extension of single-stranded DNA chains by SIEP was further verified by their binding to antibodies specific to oligonucleotides. TdTase-mediated SIEP can also be used to grow spatially defined three-dimensional DNA structures by soft lithography and is a new tool for bioinspired fabrication at the micro- and nanoscale.
Chow
Dominic C
DC
Department of Biomedical Engineering, Duke University, Durham, North Carolina 27708, USA.
Chilkoti
Ashutosh
A
D000906:Antibodies / Q000737:chemistry; D000918:Antibody Specificity; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000737:chemistry*; D003838:Deoxyadenine Nucleotides / Q000737:chemistry; D003848:Deoxyguanine Nucleotides / Q000737:chemistry; D018625:Microscopy, Atomic Force; D036103:Nanotechnology / Q000379:methods*; D009841:Oligonucleotides / Q000737:chemistry; D011089:Polydeoxyribonucleotides / Q000138:chemical synthesis*; D013052:Spectrometry, X-Ray Emission; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D013942:Thymine Nucleotides / Q000737:chemistry; D013997:Time Factors
D000906:Antibodies; D004277:DNA, Single-Stranded; D003838:Deoxyadenine Nucleotides; D003848:Deoxyguanine Nucleotides; D009841:Oligonucleotides; D011089:Polydeoxyribonucleotides; D013942:Thymine Nucleotides; C029603:deoxyguanosine triphosphate; D004253:DNA Nucleotidylexotransferase; C026600:2'-deoxyadenosine triphosphate; C024157:thymidine 5'-triphosphate