The DNA aptamers that specifically recognize ricin toxin are selected by two in vitro selection methods.
Aptamers which specifically recognize cytotoxin ricin were successfully selected using the two different in vitro selection methods. One selection method was used to isolate aptamers by affinity chromatography. Another selection method, named CE-SELEX, was carried out using CE as a separation approach. The high separation efficiency of CE evidently improved the rate of enrichment and obviously shortened the selection rounds, with near 87.2% binding just after the fourth round of selection. The aptamers A3, C1, and C5, derived from the two selection methods, were found to possess high affinity and specificity for ricin with the Kd values in the low nanomolar range, and did not recognize abrin toxin similar to ricin in the structures and properties, or BSA. Among the aptamers selected, A3 isolated by affinity chromatography shared extensive sequence similarity with C1 and C5 derived from CE-SELEX. They differed by only one base from each other. Their stable secondary structures predicted also had very similar structure motifs, and all folded a long and internal loop-embedded loop stem structure by base pairing. The ELISA and dot-blot analysis also proved that the selected DNA aptamers had the high specificity to ricin toxin.
Tang
Jijun
J
Beijing Institute of Pharmacology and Toxicology, Beijing, China.
Xie
Jianwei
J
Shao
Ningsheng
N
Yan
Yan
Y
D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000302:isolation & purification*; D001483:Base Sequence; D002846:Chromatography, Affinity; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012276:Ricin / Q000032:analysis* / Q000737:chemistry; D052156:SELEX Aptamer Technique / Q000379:methods*
D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D012276:Ricin