Difference between revisions of "Part:BBa K1653002:Experience"

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===Applications of BBa_K1653002===
 
===Applications of BBa_K1653002===
 
<p>Compare strain H1 and H2</p>
 
<p>Compare strain H1 and H2</p>
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<img class="illus" src="https://2019.igem.org/wiki/images/1/1c/T--SEFLS_Shanghai--BBa_K1653002-fig1.jpg"></img>
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[[File:T--SEFLS Shanghai--BBa K1653002-fig1.jpg]]
 
<p>Genes idi and ispA were added to the plasmid pET-YSS, yielding pET-IAY. Co-transformation of the plasmid pET-IAY with p35151 resulted in squalene production of 69.3 mg/L (strain H2), an approximately 3.7-fold increase compared to BL21(DE3) harboring pET-YSS and p35151(Figure. 2). This result indicated that the overexpression of idi and ispA increased the supply of precursor, and thus increasing the yield of squalene.&nbsp;</p>
 
<p>Genes idi and ispA were added to the plasmid pET-YSS, yielding pET-IAY. Co-transformation of the plasmid pET-IAY with p35151 resulted in squalene production of 69.3 mg/L (strain H2), an approximately 3.7-fold increase compared to BL21(DE3) harboring pET-YSS and p35151(Figure. 2). This result indicated that the overexpression of idi and ispA increased the supply of precursor, and thus increasing the yield of squalene.&nbsp;</p>
 
<img class="illus" src="https://2019.igem.org/wiki/images/0/07/T--SEFLS_Shanghai--BBa_K1653002-fig2.jpg"></img>
 
<img class="illus" src="https://2019.igem.org/wiki/images/0/07/T--SEFLS_Shanghai--BBa_K1653002-fig2.jpg"></img>
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<p class="illus-text">Figure.2 construction of strains H1, H2 &amp; H3</p>
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[[File:T--SEFLS Shanghai--BBa K1653002-fig2.jpg]]
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<p>Figure.2 construction of strains H1, H2 &amp; H3</p>
 
<p>&nbsp;Compare plasmids pETDuet-1T7-YSS-2T7-CrtN <strong>BBa_K3166998&nbsp;</strong>with<strong>&nbsp;</strong>pETDuet-rbs-idi-rbs-ispA-rbs-YSS-2T7-CrtN <strong>BBa_K3166808</strong></p>
 
<p>&nbsp;Compare plasmids pETDuet-1T7-YSS-2T7-CrtN <strong>BBa_K3166998&nbsp;</strong>with<strong>&nbsp;</strong>pETDuet-rbs-idi-rbs-ispA-rbs-YSS-2T7-CrtN <strong>BBa_K3166808</strong></p>
 
<p>By testing the pigmentation level，We discovered that the pigmentation level in the expression of YSS is higher than other strains expressing NSS KSS or thSQS, showing that YSS is relatively active, while the pigmentation level in strains containing pET-IAY-CN &nbsp;is 2.2 times that of strains expressing pET-YN, demonstrating that we can significantly increase the metabolic flux towards FPP by expressing idi and ispA.</p>
 
<p>By testing the pigmentation level，We discovered that the pigmentation level in the expression of YSS is higher than other strains expressing NSS KSS or thSQS, showing that YSS is relatively active, while the pigmentation level in strains containing pET-IAY-CN &nbsp;is 2.2 times that of strains expressing pET-YN, demonstrating that we can significantly increase the metabolic flux towards FPP by expressing idi and ispA.</p>

Revision as of 22:12, 21 October 2019


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Applications of BBa_K1653002

Compare strain H1 and H2

File:T--SEFLS Shanghai--BBa K1653002-fig1.jpg

Genes idi and ispA were added to the plasmid pET-YSS, yielding pET-IAY. Co-transformation of the plasmid pET-IAY with p35151 resulted in squalene production of 69.3 mg/L (strain H2), an approximately 3.7-fold increase compared to BL21(DE3) harboring pET-YSS and p35151(Figure. 2). This result indicated that the overexpression of idi and ispA increased the supply of precursor, and thus increasing the yield of squalene. 

<img class="illus" src="T--SEFLS_Shanghai--BBa_K1653002-fig2.jpg"></img> File:T--SEFLS Shanghai--BBa K1653002-fig2.jpg

Figure.2 construction of strains H1, H2 & H3

 Compare plasmids pETDuet-1T7-YSS-2T7-CrtN BBa_K3166998 with pETDuet-rbs-idi-rbs-ispA-rbs-YSS-2T7-CrtN BBa_K3166808

By testing the pigmentation level，We discovered that the pigmentation level in the expression of YSS is higher than other strains expressing NSS KSS or thSQS, showing that YSS is relatively active, while the pigmentation level in strains containing pET-IAY-CN  is 2.2 times that of strains expressing pET-YN, demonstrating that we can significantly increase the metabolic flux towards FPP by expressing idi and ispA.

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