Difference between revisions of "Part:BBa K3803016"

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Based on the parts (BBa_K2314913 & BBa_K2314324) from OUC_China in 2017 iGEM, we built a more efficient xylose utilization part BBa_K3803016.  
 
Based on the parts (BBa_K2314913 & BBa_K2314324) from OUC_China in 2017 iGEM, we built a more efficient xylose utilization part BBa_K3803016.  
 
Xylose reductase (XR) will first transform xylose to xylitol, and xylitol dehydrogenase (XDH) can further convert xylitol to xylulose.Then, xylulose will be converted to xylulose 5-phosphate (X5P) by the native xylulose kinase (XK). Apart from the original XR and XDH genes, considering the low copy number of native XK gene in S. cerevisiae, we introduced an extra XK gene to improve the xylose utilization ability.
 
Xylose reductase (XR) will first transform xylose to xylitol, and xylitol dehydrogenase (XDH) can further convert xylitol to xylulose.Then, xylulose will be converted to xylulose 5-phosphate (X5P) by the native xylulose kinase (XK). Apart from the original XR and XDH genes, considering the low copy number of native XK gene in S. cerevisiae, we introduced an extra XK gene to improve the xylose utilization ability.
You can see the detailed improvement here: https://2021.igem.org/wiki/images/4/41/T--Jiangnan_China--improvement-1.png
 
  
For cell growth, OUC_China achieved a final OD600 around 2.35 in YNB-based xylose media. After our improvement on their parts, we could get a final OD600 around 20. You can see the comparison of OD after improvement here: https://2021.igem.org/wiki/images/c/c7/T--Jiangnan_China--ODduibi.png
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For cell growth, OUC_China achieved a final OD600 around 2.35 in YNB-based xylose media. After our improvement on their parts, we could get a final OD600 around 20.
  
For our own product yield, our production also improved after substituting our three-gene cassette for OUC_China's two-gene one. You can see the HPLC results with different xylose utilization genes here: https://2021.igem.org/wiki/images/8/8c/T--Jiangnan_China--HPLCXYL.png
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For our own product yield, our production also improved after substituting our three-gene cassette for OUC_China's two-gene one. The improved production was very obvious after the parts improvement based on our HPLC results.
  
 
For more information about our parts and improvement, you can click here: https://2021.igem.org/Team:Jiangnan_China/Parts
 
For more information about our parts and improvement, you can click here: https://2021.igem.org/Team:Jiangnan_China/Parts

Revision as of 11:02, 16 October 2021


This is a efficient xylose utilization composite part.

Based on the parts (BBa_K2314913 & BBa_K2314324) from OUC_China in 2017 iGEM, we built a more efficient xylose utilization part BBa_K3803016. Xylose reductase (XR) will first transform xylose to xylitol, and xylitol dehydrogenase (XDH) can further convert xylitol to xylulose.Then, xylulose will be converted to xylulose 5-phosphate (X5P) by the native xylulose kinase (XK). Apart from the original XR and XDH genes, considering the low copy number of native XK gene in S. cerevisiae, we introduced an extra XK gene to improve the xylose utilization ability.

For cell growth, OUC_China achieved a final OD600 around 2.35 in YNB-based xylose media. After our improvement on their parts, we could get a final OD600 around 20.

For our own product yield, our production also improved after substituting our three-gene cassette for OUC_China's two-gene one. The improved production was very obvious after the parts improvement based on our HPLC results.

For more information about our parts and improvement, you can click here: https://2021.igem.org/Team:Jiangnan_China/Parts

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 805
    Illegal XbaI site found at 6464
    Illegal PstI site found at 2114
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 805
    Illegal PstI site found at 2114
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 805
    Illegal BglII site found at 1265
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 805
    Illegal XbaI site found at 6464
    Illegal PstI site found at 2114
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 805
    Illegal XbaI site found at 6464
    Illegal PstI site found at 2114
    Illegal AgeI site found at 1936
  • 1000
    COMPATIBLE WITH RFC[1000]