Team:USTC/Notebook/10 1
Notebook
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Date: 10.7
Recorder: Kaiyue Ma The results of the observation through the fluorescence microscope are showed below.
The yeasts with plasmid YeLGAP carrying the part of pGAL+GFP:
The intensity of the fluorescence is relatively weak as expected. Because of the absence of AD and BD, the expression of the gene under the regulation of pGAL is shut off. Next, we'll construct the yeasts containing all circuits of our Propri-ontein system.
The yeasts with plasmid YeLGAP carrying the part of pGAL+GFP and plasmid YeWGAP carrying the part of SUP35+BD:
The yeasts with plasmid YeWGAP carrying the part of sfGFP1-10:
The yeasts with plasmid YeWGAP carrying the part of sfGFP1-10 and plasmid YeUGAP carrying the part of SUP35+sfGFP11:
Comparing the two pictures above, it's apparent that there are intensive green spots inside the yeasts in the second picture, but none in the first. We assumed that the green spots are caused by the combination of sfGFP11 and sfGFP1-10, and the light green of the yeasts are caused by sfGFP1-10 itself. The same phenomena were observed several times. In addition, when the PSB1C3 carrying the part of sfGFP1-10 and the PSB1A3 carrying the part of sfGFP11 ware expressed together in E.Coli, it doesn't present stonger fluorescence intensity than either plasmid is expressed in E.Coli respectively. We assume that different metabolic stress of two plamids causes inditinct results. Because plasmid YeUGAP carrying sup35 besides fluorescence intensity, the approximate DNA length may decrease metabolic stress difference between two kinds of plasmids. The results of heat shock can also prove our assumption. Under same expression level, expression of sup35, which leads to plasmids YeUGAP aggregate, will apparently decrease the number of intensive green spot.
Date: 10.8
Recorder: Xuefeng Meng and Chengle Zhang
The results of the observation through the fluorescence microscope are showed below.
The yeasts with plasmid YeWGAP carrying the part of sfGFP1-10 and plasmid YeUGAP carrying the part of SUP35+sfGFP11 after heat shock of 42 degree Celsius for 1 hour:
The yeasts with plasmid YeWGAP carrying the part of sfGFP1-10 and plasmid YeUGAP carrying the part of SUP35+sfGFP11 after heat shock of 42 degree Celsius for 2 hours:
The yeasts with plasmid YeWGAP carrying the part of sfGFP1-10 and plasmid YeUGAP carrying the part of SUP35+sfGFP11 after heat shock of 37 degree Celsius for 2 hours:
Date:10.9 Recorder: Meng Xuefeng Green fluorescence detecting of pro-priontein
Figure 1: 30℃ 1h 0mM GdnHCl
Figure 2: 42℃ 1h 0.25mM GdnHCl
Figure 3: 42℃ 1h 0.5mM GdnHCl
Figure 4: 42℃ 1h 3mM GdnHCl
Figure 5: 42℃ 1h 5mM GdnHCl
