Difference between revisions of "Part:BBa K3166059:Experience"
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===Applications of BBa_K3166059=== | ===Applications of BBa_K3166059=== | ||
| + | <p>The genes dxs and ispG were placed on the backbone of low-copy plasmid pBBR1MCS-2, and the promoter was medium strength lac promoters, to construct plasmid pBBR1MCS-2-dxs-ispG (pMEP-DG).</p> | ||
| + | <p>Genes ispH, idi, and ispA were placed on the backbone of high-copy plasmid pETDuet-1, and the promoter was strong promoter T7 to construct plasmid pETDuet-1-1T7-IspH-Idi-IspA-Yss (pET-HIAY).</p> | ||
| + | <p>When Dxs and IspG were overexpressed, the squalene yield increased by 4.3 times. We speculated that the overexpressed gene ispG was constructed on low-copy plasmids and was controlled by promoters of medium strength, so it didn’t produce large amounts of harmful intermediate HMBPP. And overexpressing Dxs promoted the metabolic flow, increasing the squalene yield.</p> | ||
| + | https://2019.igem.org/wiki/images/9/91/T--SEFLS_Shanghai--BBa_K1088004-fig1.png | ||
| + | <p>H3: p35151/pET-HIAY</p> | ||
| + | <p>H5: p35151/pMEP-DG/pET-HIAY</p> | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 22:23, 21 October 2019
This experience page is provided so that any user may enter their experience using this part.
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how you used this part and how it worked out.
Applications of BBa_K3166059
The genes dxs and ispG were placed on the backbone of low-copy plasmid pBBR1MCS-2, and the promoter was medium strength lac promoters, to construct plasmid pBBR1MCS-2-dxs-ispG (pMEP-DG).
Genes ispH, idi, and ispA were placed on the backbone of high-copy plasmid pETDuet-1, and the promoter was strong promoter T7 to construct plasmid pETDuet-1-1T7-IspH-Idi-IspA-Yss (pET-HIAY).
When Dxs and IspG were overexpressed, the squalene yield increased by 4.3 times. We speculated that the overexpressed gene ispG was constructed on low-copy plasmids and was controlled by promoters of medium strength, so it didn’t produce large amounts of harmful intermediate HMBPP. And overexpressing Dxs promoted the metabolic flow, increasing the squalene yield.
H3: p35151/pET-HIAY
H5: p35151/pMEP-DG/pET-HIAY
User Reviews
UNIQ9c351e2a49e1f230-partinfo-00000000-QINU UNIQ9c351e2a49e1f230-partinfo-00000001-QINU