Object: Rana grylio virus stress Rana dybowskii, the genome RNA were extracted from Rana skin,sequenced the Toll like receptor (TLR) genes, analysis TLRs gene temporal and spatial expression pattern, large number TLRs are synchronized expression as well as each other, whether they are functionally related.Method:Extract total RNA Collect from Rana dybowskii skin tissue.RT-PCR amplification with a pair of degenerate primers designed according to the terminal conserved sequences of TLR precursors of genus Rana, Fluctuated changes of TLR mRNA expression at each time are showed below.TLRl expression level was raised rapidly to the peak at 1st hour.After one reversible reduction, it went back to peak at 6tb hour.Then was the down-regulation and remained lower than the control group.TLR2 expression level stayed lower than the control group within 2 days.It started to up-regulate at 12th hour and reached the peak at 3rd day, while it went lower than the control group after 4th day.TLR4 expression level performed down-regulation within 2 hours.It started to go up at 2nd hour, then reached the peak at 6th hour.After 12 hours, it stayed lower than the control group all the time.TLR5 expression was down-regulated within 2 hours.Then it started to raise at 2nd hour, and reached the peak at 3rd hour.After a slow reversible down-regulation, it raised to the peak again at 4th day.After 5 days, it stayed lower than the control group all the time.TLR8 expression level was raised to the peak rapidly at 1st hour.After a reversible up-regulation, it went back to the peak rapidly at 3rd hour.Then it was slowly down-regulated to the level that lower than the control group.Conclusion: It is demonstrated that all the peptides showed a significant anti-Rana grylio virus activity, and with the increase in gene expression, the virus was gradually cleared.Ranatuerin-2YJ,in particular, also showed a broad-spectrum antibacterial activity against the Gram-positive and Gram-negative bacteria among all the peptides with a lower hemolysis activity.TLRs were not only involved in innate immune response caused by RGV, but also launched the acquired immune response.They were extremely important to the body on resisting viruses.To avoid competing with MyD88 molecules, each TLR were regulated to ensure balance.When stressed by virus, TLR1 and TLR8 were up-regulated immediately to the peak, and the differences between before and after the peak was the most obvious, indicating that the Ranatuerin-2YJ, TLR1 and TLR8 in vivo plays an important role in the protection against pathogens invasion.