Optimization and application of prokaryotic expression of dsRNA-StPPO fermentation process
RNA interference(RNAi)is a phenomenon of highly efficient and specific degradation of homologous mRNA induced by double-stranded RNA(dsRNA),which triggers specific cellular inhibition or silencing of homologous target genes.RNAi technology has been applied in many fields such as pest control,bacterial inhibition,antiviral,etc.,with less application in controlling the quality dete-rioration of agricultural products.To optimize the expression and fermentation conditions of dsRNA for the gene encoding potato polyphenol oxidase StPPO,Escherichia coli HT115(DE3)RNase Ⅲ deficient strain was employed as the fermentation platform and recombinant vec-tor was constructed using the L4440 vector as the backbone.Furthermore,the biological function of dsRNA-StPPO was validated.The re-sults showed that the yield of the target dsRNA was the highest at 3.95 µg/mL at the concentration of isopropylthiogalactoside(IPTG)in-ducer at 0.6 mmol/L,induction time of 6 h,plus yeast extract of 0.48％,temperature of 39℃,bottling volume of 45 mL/250 mL,and rotation speed of 189 r/min.After immersion in dsRNA-StPPO solution,fresh-cut potatoes in the treatment group had higher L*,lower a* and browning index,and lower polyphenol oxidase enzyme activity in the tissues compared with the control group,suggesting that the application of dsRNA-StPPO can inhibit the expression of the StPPO gene and effectively control the browning phenomenon of fresh-cut potato.
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维普
