Part:BBa_K4340601
PYL8
PYL8 is a highly efficient ABA inhibitor. It binds to ABA and stops the signaling of ABA. We next tested whether the hormone binding domain can inhibit and attenuate the effect of ABA (plant hormone that inhibits germination and plant growth) to plant germination. The pET11a and PYL8 E.coli culture groups, and a group with ABA treatment and PYL8 E. coli culture had a higher germinated percentage than the group without ABA (blank). This indicated that the PYL8 had a positive effect on the seed germination stage both with ABA and without ABA. (Figure 1) In the PYL8 group results, the percentage of germinated soybeans with PYL8 E.coli culture and ABA was significantly higher than in groups only with ABA. This shows that the PYL8 worked to produce an inhibitor of ABA. Notably, the PYL8 group (only added PYL8 cell culture) is even higher than the group without ABA. This demonstrated that PYL8 can discharge the effect of ABA, and even benefit seed germination squarely. (Figure 2)
Experiment 1: Hormone-treated germination test
To sum up, the efficiency of the plasmids ranked is PYL8> NSP4-T2R4 (slightly higher than) pET11a.
Experiment 2: Western Blot and Real-time PCR
We have conducted a western blot experiment to validate the quality of protein expression of PYL8 and NSP4-T2R4. In the experiment, there is a clear band of PYL8 in both 10ul and 20ul samples at 35 kDa. (Figure 3)
In the real-time quantitative PCR test, we found the RNA expression of both genes, indicating that the PYL8 expressed RNA successfully. (Figure 4)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 305
- 1000COMPATIBLE WITH RFC[1000]
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