Difference between revisions of "Part:BBa K3402055"

 
 
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<partinfo>BBa_K3402055 short</partinfo>
 
<partinfo>BBa_K3402055 short</partinfo>
  
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This device is composed of 50bp-up<i>PXA1</i>(BBa_K3402037), Six site(BBa_K3402032), P<i>galk</i>(BBa_K3402033), <i>Rec</i>(BBa_K3402034), T<i>gki</i>(BBa_K3402019), <i>hph</i>(BBa_K3402012), P<i>tef1</i>(BBa_K3402007), <i>UGTB</i>(BBa_K3402011), T<i>syn7</i>(BBa_K3402001), 50bp-do<i>PXA1</i>(BBa_K3402038).
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This device is composed of 50bp-up<i>PXA1</i>([https://parts.igem.org/Part:BBa_K3402037# BBa_K3402037]), Six site([https://parts.igem.org/Part:BBa_K3402032# BBa_K3402032]), P<i>galk</i>([https://parts.igem.org/Part:BBa_K3402033# BBa_K3402033]), <i>Rec</i>([https://parts.igem.org/Part:BBa_K3402034# BBa_K3402034]), T<i>gki</i>([https://parts.igem.org/Part:BBa_K3402019# BBa_K3402019]), <i>hph</i>([https://parts.igem.org/Part:BBa_K3402012# BBa_K3402012]), P<i>tef1</i>([https://parts.igem.org/Part:BBa_K3402007# BBa_K3402007]), <i>UGTB</i>([https://parts.igem.org/Part:BBa_K3402011# BBa_K3402011]), T<i>syn7</i>([https://parts.igem.org/Part:BBa_K3402001# BBa_K3402001]), 50bp-do<i>PXA1</i>([https://parts.igem.org/Part:BBa_K3402038# BBa_K3402038]).
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[[Image:Over-expression UGTB cassette.png|700px|]]
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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up<i>PXA1</i> and do<i>PXA1</i> are homologous arms, which means this device will edit the <i>PXA1</i> site. The β-Rec/six self-excising system will help to excise the hygromycin resistance gene. UDP-glucosyltransferase B (<i>UGTB</i>) is the key gene in the synthesis of sophorolipids.
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<br>
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Choose <i>PXA1</i> as the gene editing site. Three strong promoters P<i>tef1</i>, P<i>eno</i> and P<i>gki</i> were knocked in <i>PXA1</i> site to over-express UDP-glucosyltransferase B (<i>UGTB</i>).
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<br>
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Then we transformed the corresponding over-expression fragments and the Cas9 and sgRNA expression cassette to the wild-type <i>Starmerella bombicola</i>.
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<br>
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After incubation and fermentation, test the yield and acid/lactone ratio of sophorolipids produced by different strains.
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After over-expressing <i>UGTB</i>, the yield could be increased a lot than that of the control.
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[[Image:Table-The yield of sophorolipids produced by different strains.jpeg|600px|thumb|center|'''Table. 1''' The yield of sophorolipids produced by different strains]]
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[[Image:Chart-The yield of sophorolipids by over-expression of UGTB.png|500px|thumb|center|'''Chart.1''' The yield of sophorolipids by over-expression of UGTB
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<br>
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('''W: '''wild type; '''S1: '''::Ptef1-UGTB; '''S2: '''::Peno-UGTB; '''S3: '''::Pgki-UGTB)]]
  
 
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Latest revision as of 18:11, 27 October 2020


Over-expression UGTB cassette

This device is composed of 50bp-upPXA1(BBa_K3402037), Six site(BBa_K3402032), Pgalk(BBa_K3402033), Rec(BBa_K3402034), Tgki(BBa_K3402019), hph(BBa_K3402012), Ptef1(BBa_K3402007), UGTB(BBa_K3402011), Tsyn7(BBa_K3402001), 50bp-doPXA1(BBa_K3402038).

Over-expression UGTB cassette.png

Usage and Biology

upPXA1 and doPXA1 are homologous arms, which means this device will edit the PXA1 site. The β-Rec/six self-excising system will help to excise the hygromycin resistance gene. UDP-glucosyltransferase B (UGTB) is the key gene in the synthesis of sophorolipids.
Choose PXA1 as the gene editing site. Three strong promoters Ptef1, Peno and Pgki were knocked in PXA1 site to over-express UDP-glucosyltransferase B (UGTB).
Then we transformed the corresponding over-expression fragments and the Cas9 and sgRNA expression cassette to the wild-type Starmerella bombicola.
After incubation and fermentation, test the yield and acid/lactone ratio of sophorolipids produced by different strains. After over-expressing UGTB, the yield could be increased a lot than that of the control.

Table. 1 The yield of sophorolipids produced by different strains
Chart.1 The yield of sophorolipids by over-expression of UGTB
(W: wild type; S1: ::Ptef1-UGTB; S2: ::Peno-UGTB; S3: ::Pgki-UGTB)

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1076
    Illegal BglII site found at 5893
    Illegal BamHI site found at 3479
    Illegal XhoI site found at 1910
    Illegal XhoI site found at 4631
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 219
    Illegal AgeI site found at 5315
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 5474
    Illegal BsaI.rc site found at 5429