A nano-circular single-stranded DNA as a novel tool for SNPs detection.
Analysis of single nucleotides polymorphisms (SNPs) is very important for the elucidation of gene-based physiological differences. For high-throughput applications, detection systems need to be highly selective, highly sensitive, and simple. In this study, we investigated whether synchronous transcription and translation from nano-circular single-stranded DNA (ssDNA) might be useful for the detection of SNPs. A nano-circular probe ssDNA was designed to contain codons for a (His)6 peptide, and the sense mRNA transcribed from this ssDNA contains a Shine-Dalgarno sequence, a start AUG codon, 6 His codons (CAU), and a stop UAG codon. The entire circular ssDNA is 55 nt and contains T20 as a linker sequence and a binding site for the target SNP-containing DNAs. Our results that show SNPs can be detected by the cell-free synthesis of the (His)6 peptide from this ssDNA. Because this method allows sequence distinction, signal amplification, and easy detection in one system, it should improve the efficacy of high-throughput gene analysis.
Ohmichi
Tatsuo
T
Frontier Institute for Biomolecular Engineering Research (FIBER), Konan University, Higashinada-ku, Kobe, Japan.
Takashima
Aki
A
Sugimoto
Naoki
N
D020137:Base Pair Mismatch; D001483:Base Sequence; D001665:Binding Sites; D002474:Cell-Free System; D003062:Codon; D018387:Codon, Initiator; D018388:Codon, Terminator; D004270:DNA, Circular / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry*; D005821:Genetic Techniques; D006639:Histidine / Q000737:chemistry; D008956:Models, Chemical; D008969:Molecular Sequence Data; D049329:Nanostructures / Q000737:chemistry*; D036103:Nanotechnology / Q000379:methods*; D009693:Nucleic Acid Hybridization; D010455:Peptides / Q000737:chemistry; D020641:Polymorphism, Single Nucleotide; D014176:Protein Biosynthesis; D017422:Sequence Analysis, DNA / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014158:Transcription, Genetic
D003062:Codon; D018387:Codon, Initiator; D018388:Codon, Terminator; D004270:DNA, Circular; D004277:DNA, Single-Stranded; D010455:Peptides; D006639:Histidine