Abstract:[Background] Resistance gene is critical to molecular biology research, however, containing the massive size always restricts its usage like limiting the length of the target sequence inserted into the vector. [Objective] Exploring resistance gene similar function of RNA aptamer simple molecules to optimize capacity of the vector and enlarge the target gene expressing section. [Methods] The tRNA scaffold recombinant RNA expressing strategy was used to enrich ribonucleic acid aptamer of antibiotic in vivo. Screening and verifying resistance of the RNA aptamer of the aminoglycosides antibiotic neomycin B. [Results] The tRNA scaffold recombinant RNA expressing vector was constructed and successfully applied to select in vivo. The two RNA aptamers, A Site and Avirus, were proved that could resist neomycin B with 19 μg/mL on LB plate and 30 μg/mL in liquid culture. [Conclusion] RNA aptamer could bind with antibiotic, leading to being a new kind of resistance gene. This strategy is expected to optimize the resistance gene, reduce plasmid vector capacity, which provides a new method for molecular biology research.
