RNA适配体作为类抗性基因的可行性

【背景】抗性基因(ResistanceGene)在分子生物学研究中具有重要作用,但其基因片段较大,限制了载体中插入目的序列的长度。【目的】探究分子量较小的RNA适配体的类抗性基因作用,用于优化载体载量和扩展目的基因片段。【方法】基于tRNA支架在体内富集表达RNA适配体,筛选验证氨基糖苷类抗生素新霉素B的RNA适配体的抗性作用。【结果】构建tRNA支架重组RNA表达载体并进行体内筛选,得到A Site和Avirus这2个RNA适配体分子,能够耐受新霉素B浓度为19μg/mL(固体培养基)和30μg/mL(液体培养基)。【结论】RNA适配体可发挥类抗性基因的功能,本文策略有望用于抗性基因的优化,缩小质粒载体载量,为分子克隆技术提供新的手段。

A feasibility study into ribonucleic acid aptamer being antibiotic resistance gene

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 19μg/mL on LB plate and 30μg/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.