Team:FAFU-CHINA/Results
Result
Preliminatry experiment.
The effect of different homologous dsRNA of CSBV to the expression of CSBV’s mRNA.
To find the most effective homologous dsRNA of CSBV for our project, we tested the pupation rate of honeybee larvae under different dsRNA. We added different homologous dsRNA of CSBV (dsHelicase, dsProtease, dsRdRp and dsVP1) into forage and fed to the infected larvae. After 12 hours, We fed the larvae with extracted fluid of CSBV, and kept feeding it in the following day.
According to the statistics we collected, infected larvae fed with dsRdRp have the highest pupation rate among all the experimental groups.
The effect of CSBV·Silencer in curing the disease.
We fed the infected hives with our CSBV·Silencer, and collected the following statistics under the help of professional beekeepers from College of Bee Science.
1.The decrease of the number of infected larvae.
Compared with the control group, it is quite obvious that the infected larvae in medium concentration group and in high concentration group dropped dramatically by 91.44 percent and 94 87.91 percent after 14 days while the infected larvae in control group showed 65.82 percent increase after 7 days.
2.The effect of dsRdRp on the number of sealed brood.
The number of sealed brood is another indicator of the effect of CSBV.It is an necessary stage of larval pupation. In this graph, the number of sealed brood in medium concentration and high concentration increases by about 23.68% and 30.10% after 21days. Similarly, the result witnesses a significant improvement in the pupation rate.
3.The increasing population.
We counted the number adult honeybees to study the changes of population.The changes are just as you see. For the high concentration group alone, the growth of population had reached almost 60% in just 21 days. On the contrary, the control group showed a 27.78% decrease after 21 days.
These statistics directly proved that, our product, CSBV.Silencer 1.0 does have a remarkable effect in the treatment to Sacbrood disease.
The effect of galactose to induce GFP expression at different concentrations and different time.
1.The galactose at three different concentrations can induce GFP expression efficiently and have little influence on the level of GFP
We transformed GFP-pYES2 into the yeast and let the recombinant strain grow to mid-log phase in YEPD medium containing 2% glucose. And then we transferred it to medium, containing 2%, 0.1% or 0.02% galactose, and assayed for GFP fluorescence.
According to the statistics we collected, we find that the production of GFP protein is quite high even though at 0.02% galactose and with the increase of its concentration, the level of GFP have no remarkable change.
1.GFP production sharply increases and a peak shows up in about 24h under the induction of galactose.
The yeast transformants were grown in YEPD medium. At a cell density of 2 ×107 ml-1,
cultures were washed and galactose added to a final concentration of 2% and the fluorescence value of samples was measured every 5h.
As the diagram shown, we can find that GFP production sharply increases in 10h and a peak shows up in about 24h and then it decreases gradually.
In conclusion, we can find using 0.02% galactose to induce the expression of GFP in 24h is a reasonable and efficient way to get the most production of GFP protein. It means that we can induce the expression of T7 RNAP gene efficiently under the same condition.
