The PLA Plasmid Of Recombinant Lactobacillus QPCR Method For Detection Of Different Conditions Of PCN

Lactic acid bacteria belongs to probiotics, which has obvious probiotic role and takes orallyconveniently. So the researches that use lactic acid bacteria as oral vaccine are increasing inrecent years. As a kind of the new lactobacillus plasmid expression system that is builtsuccessfully, protein expression index is the basis of the further study.In order to build a rapid, accurate and specific quantitative testing method of expression ofrecombinant plasmid copy number, we make analysis on recombinant plasmid copy numberaccording to SYBR Green I principle of fluorescence quantitative analysis. Assessinglactobacillus genome and recombinant plasmid mediated DNA by computer software Oligo5.Designing specific primers for good and building qPCR detection method for absolutequantification. By using the method, determining recombinant plasmid copy number. The resultsshow that real time fluorescence quantitative PCR detection method could confirm recombinantplasmid copy number rapidly, accurately, specifically and sensitively. Lactobacillus plasmidcopy number can still exist stably and keep the high number type property in the non-resistancetraining conditions and this result provides the secure basis for the further research about therelevance of the plasmid copy number and expression level in the process of fermentation.To ensure recombinant expression vector whether to keep probiotic properties of theoriginal or not, its colonization and sustained expression in intestinal is an important index.Digestion test in vitro for recombinant Lactobacillus is carried out by the simulated digestionexperiment method in common use. The experiments show that recombinant lactobacillus cankeep the original growth, expression characteristics. The results provides secure basis for furtherapplication in the entity animal...