DNA detection system using molecularly imprinted polymer as the gel matrix in electrophoresis.
To develop a simple and inexpensive method for DNA detection, we prepared a molecularly imprinted polymer (MIP) for recognizing a specific double-stranded DNA (dsDNA) sequence and used it in an electrophoretic gel matrix. The MIP gel has many binding sites that are complementary in size, shape, and arrangement of functional groups of the target dsDNA sequence. During MIP gel electrophoresis (MIPGE), migration of the target dsDNA should be hindered by the capture effect of the binding sites in the MIP gel. This was confirmed by observation of deviations from the linear relationship between the migration distances of the DNA standard size markers in the polyacrylamide gel and those in the MIP gel. The migration distances of nontarget dsDNA maintained a linear relationship, however. In addition, the sequence selectivity of dsDNA in this method was investigated by using the Ha-ras gene and its point mutants. Except for A.T to T.A base pair substitution, mutant dsDNA (for example, substitution from A.T to C.G and from G.C to T.A) could be distinguished from the target (wild-type) dsDNA. Although some improvement in A.T (T.A) base pair distinction is still needed, this study is the first to demonstrate detection of a specific dsDNA sequence with MIPs and, as such, opens up a new realm for practical applications of MIPs.
Ogiso
Masayo
M
Research Center of Advanced Bionics, National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562, Japan.
Minoura
Norihiko
N
Shinbo
Toshio
T
Shimizu
Toshimi
T
D004247:DNA / Q000032:analysis* / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D014227:Triazines; D014753:Vinyl Compounds
C500826:2-vinyl-4,6-diamino-1,3,5-triazine; D014227:Triazines; D014753:Vinyl Compounds; D004247:DNA