The Establishment Of Screening Model For Artificial Trans-encoded RNAs In Bacteria

As the study progresses, in recent years scientists have gradually discovering and understanding the widespread presence of a new class of RNA molecules in living organisms, that is, small non-coding RNA (small non-coding RNA), their size usually less than 500bp, cannot be translated into protein, but have a wide impact on the regulation of organism life activity. At present, we also found in eukaryotes there were some sRNAs such as microRNA and siRNA and many other small non-coding RNA molecules.?Along with the deepening of the study, people in prokaryotic organisms (especially bacteria) were found in eukaryotic small RNA-like a very high degree of RNA molecules, named for trans-coding small RNA (trans-encoded sRNA); they are by and features gene complementary pairs to reduce the level of translation of target genes and affect its stability. RNA interference (RNAi) technology has developed into a study on the expression regulation in eukaryotic organisms of the principal means. To date, homologous recombination, gene knockout technology remains the main tool for study of gene expression in prokaryotes. In prokaryotes, establishing a similar to RNAi gene silencing technology with a wide range of opportunities for development.At the early stage of research, we use bio-informatics related software, analysis of some endogenous anti-bacterial effect of small primary and secondary structure of RNA, the bacteria was first introduced by trans-acting principle design of small non-coding RNA. According to this principle, for endogenous and exogenous gene of Escherichia coli we designed a series of atsRNA, to turn them into host cells, by detecting expression of target genes, we found the most atsRNA to varying degrees of silent expression of target genes, of which about interference with the highest efficiency of up to 70%. We have created a new method to study gene function in bacteria.In order to further implement these new methods of application, how to achieve a certain target gene screening of a series of atsRNA, convenient and intuitive to interference of high efficiency atsRNA identified, we conducted a new study. In Escherichia coli in the construction has double mass grain filter model, select has glmS and ompR as target genes, under the designing principles, respectively design has G and R series of atsRNA, by determination and target gene fusion of report gene lacZ of activity, various atsRNA for target gene rendering out has different interference level, which highest reached 67% above.Through experimental design we get artificial trans small non-coding RNA in gene silencing of gram-negative bacteria played a prominent role, to explore whether the design principles in setting for Gram-positive bacteria, we designed a series of virulence gene of Staphylococcus aureus for atsRNA, but did not find their target genes have a noticeable effect.To sum up, we have created a atsRNA filter to implement a quick and effective way, by double-plasmid screening model, simple visual comparison of different atsRNA jamming efficiency, determination and the adoption of the report gene activity, indirectly reflects the performance of easy detection of target gene expression. atsRNA screening model of effect in small non-coding RNA is important extension and application of design principles, has better prospects, can make new contributions for the treatment of bacterial infections.