Team:Shanghai City United/Results
Results
Figure 1. electrophoregram of XynA
Figure 1 is about the electrolysis of XynA. The result of electrolysis can clearly
show whether the XynA is successfully amplified. XynA was amplified by PCR. The length of XynA is 650 bp. By
compared the marker and the place of DNA, the PCR of XynA is successful.
Figure 2. electrophoregram of pMD19-T-xynA enzyme-digested product
Channel 1: pMD19-T-xynA-LCX-ZZY, correct
Channel 2: pMD19-T-xynA-HHJ, correct
Channel 3: pMD19-T-xynA-HX, incorrect
Channel 4: pMD19-T-xynA-LJL, incorrect
Channel 5: pMD19-T-xynA-0, incorrect
Channel 6: pMD19-T-xynA-SXY, incorrect
Channel 7: pMD19-T-xynA-CYF, incorrect
Figure 3. electrophoregram of enzyme-digested products of pSIP403 and PUS
Channel 1~2: pSIP403, 6kbp, correct
Channel 3~4: PUS, 300bp, correct
Figure 4. SDS PAGE result of BL21(DE3)/pMD19-xynA, Coomassie blue staining.
Figure 5. Protein molecular weight result of XynA by Snapgene
After the sample has been centrifuged and sonicated. we put it to
SDS-PAGE(SDS-polyacrylamide gel
electrophoresis) which could determine the level of protein expression. As seen in the gel map (Maker,
culture solution, intracellular supernatant, intracellular precipitation), the second red line of Marker
represents 25KDa and the target protein should be 23.28kDa(Fig. 5). The blue line in the culture solution
was near 23 KDa, it indicates that our protein could be sucessfully expressed in E. coli.
Figure 6. sequencing result of PMD19-T-XynA
According to the sequencing results (Fig.6) from the sequencing company, the solid
black line is the ideal
state of PMD19-T-XynA and the DNA sequence of our plasmid matches the profile.
Figure 7. DNS test results of E. coli/pMD19-xynA
The enzyme activity test results showed groups from the left to the right,
respectively: blank, culture medium supernatant * 2, intracellular supernatant * 2, and intracellular
precipitation * 2. It could be seen by naked eyes that in the seven bottles of solution, the culture medium
solution showed red after DNS reaction, which indicated that we had xylanase with well enzyme activity to
degrade xylan and this result is consistent with the SDS PAGE result.
Figure 8. Colony PCR results of Lactobacillus reuteri/pSIP403-PUS-xynA and marker maps
In Figure 8, it can be clearly seen that our plasmid pSIP403-PUS-xynA has been
successfully transformed
into Lactobacillus reuteri.
Future Plan
1. Conduct the enzyme activity tests of Lactobacillus reuteri/pSIP403-PUS-xynA to
analyze the performance in Lactobacillus reuteri.
2. Design control tests to determine the optimal conditions of XynA protein
expression in E. coli
3. Explore the safety and stability of the protein