Selection of ssDNA Aptamers of Listeria monocytogenes by Whole SELEX
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Abstract:
In this paper, aptamers against Listeria monocytogenes live cells were screened by systematic evolution of ligands by exponential enrichment (SELEX) from single-tranded DNA (ssDNA) library which contained 40 random bases. During the SELEX selection, affinity ability between ssDNA sub-library and L. monocytogenes was evaluated by enzyme linked aptamer absorbent assay (ELASA). The characteristics of aptamers including sequence, secondary structure, affinity and specificity to L. monocytogenes were studied. The results revealed that aptamers were enriched after 15 SELEX rounds and the affinity against target enhannced round by round. The aptamers of 23 different sequences obtained via sequencing were divided into 3 groups, according to the similarity of secondary structure predicted by RNA structure program. Also, it indicated that the secondary structure was related with the binding ability against the target. Two aptamers named Lma-21 and Lma-35 with high affinity could specifically recognize L. monocytogenes. This work provided necessary materials for the development of new type of detection reagents against L. monocytogenes.
