The Model Of Rapid Aging By The Ctl-2 Gene RNA Interference In Carenorhabditis Elegans

C.elegans Peroxisomal catalase, CTL-2 is a sort of main catalase inCaenorhabditis elegans in vivo. It has effect of scavenging of free radicals,and repair of radical-induced damage of biomacromolecules. Someoneusing genetic mutants that lack of peroxisomal catalase CTL-2 causes aprogeric phenotype in C. elegans. Lack of peroxisomal catalase also affectsthe developmental program of C. elegans, since ctl-2 mutants' exhibitdecreased egg laying capacity. 'We think ctl-2 gene relate to oxidativedamage and its induced aging. Studies of ctl-2 gene function will redound tothe study of aging mechanism and anti-oxidative drug activity.RNA interference technology can separated especial gene silence inCaenorhabditis elegans in vivo and in vitro, that result in function lose orreduce mutation. The phenotypes of inhibition of the gene are similar to thatby direct deletion of the same gene. RNA interference technology becomesa means of research gene function.Here we use the nematode Caenorhabditis elegans (C.elegans) as amodel system to investigate the function of the peroxisomal catalase.Based on the technique of double-stranded RNA interference (RNAi), weestablished the model of rapid aging in C.elegans by controlling theexpression of ctl-2 gene. We observe the change of C.elegans after RNAinterference and the effect of gene silence. To make certain that relativity ofthe ctl-2 mRNA expression, life-span and egg laying capacity, we amplifythe ctl-2 gene by the method of real time RT-PCR.Based on the technique of gene engineering technology, wesuccessfully established the pPD129.36-ctl-2 vector and E.coli HT115(DE3),that can produced dsRNA by IPTG induced. We explored feedingas a means of delivering dsRNA for RNAi. From the change of the ctl-2mRNA expression, life-span and egg laying capacity, we gained this resultand conclusion:In this assay, we survey ctl-2 mRNA expression in different culturetemperature and IPTG concentration. At low temperature, RNA interferencetype C. elegans ctl-2 mRNA increase several fold compare with thewildness type. But, the two type C. elegans at different temperature have nodifference in the RNA interference type. It may be the ctl-2 mRNAdegradation in the RNA interference type. The result also confirms theconclusion that IPTG induce dsRNA expression is successful.Another, to determine the optimal concentration of IPTG for producingRNAi phenotypes, we chose the quantity of eggs and longevity ofC.elegans as the target. As a result, we found that 1uM IPTG gave us thehighest penetrance of phenotypes. The data analytic result confirms theconclusion that there is consanguineous relation between ctl-2 gene andage-related gene, and there is a free back between ctl-2 mRNA andlife-span. It may be appeared to scavenging system of free radicals oroxidative damage system.Except upwards phenomenon, we observed thatctl-2 gene related to growth differentiation system.In this study, our work establish experiment base for discussing agingmechanism. We think that RNAi C.elegans could as a model system toinvestigate the effect of anti-oxidative drug and the aging mechanism.