科研之友
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摘要
A competitive inhibition method based on a molecular beacon containing 20 nucleotides was developed for the active site analysis and modification of two organophosphorusate pesticides aptamers that had been selected in our previous study. The results indicated that the designed molecular beacon formed a hairpin structure,and the loop could be opened up in the presence of complementary aptamer sequence. The ratio of molecular beacon to aptamer of 1.25 to 1, incubated time of 50 min and incubation at room temperature were chosen as optimal conditions of active site analysis. Under optimal conditions,the Loop2-4 was a mutual active site for four organophosphorus pesticides, Loop2-3 and the nucleotides at 3' and 5' of the SS4-54 aptamer were important active sites for phorate, Loop2-2 and Loop4-2 were mutual active sites for profenofos and isocarbophos, Loop4-3 was a mutual active site for profenofos and omethoate, Loop2-1 and Loop4-1 were important active sites for isocarbophos. The binding activity for profenofos and isocarbophos of SS24-PJ-35 aptamers modified by gene splicing significantly increased.
