Difference between revisions of "Part:BBa K5271011"
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<partinfo>BBa_K5271011 short</partinfo> | <partinfo>BBa_K5271011 short</partinfo> | ||
| − | A nanobody composed of two binding regions of EGFR and HER2 linked by a fragment of Fc region of the heavy chain of human IgG | + | A nanobody composed of two binding regions of EGFR and HER2 linked by a fragment of Fc region of the heavy chain of human IgG. |
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| + | =Profile= | ||
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| + | *Name: HER2nb-Fc-EGFRnb | ||
| + | *Base Pairs: 1515 bp | ||
| + | *Amino acid: 527 a.a | ||
| + | *Origin: Synthetic | ||
| + | *Property: A dual targeting nanobody with a molecular weight of 55 kDa consisting of a Fc fragment and two HER2 and EGFR nanobody binding domains that specifically bind to EGFR and HER2. | ||
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| + | =Usage and Biology= | ||
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| + | =Expression= | ||
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| + | :::https://static.igem.wiki/teams/5271/part-figures/sds-gel-her2-fc-egfr-16c-30c.png | ||
| + | :::'''Figure 2. Protein expression of HER2-FcC-EGFR.''' Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Total 30℃ HER2-FcC-EGFR in 6hr uninduced fraction; lane 3, Total 30℃ HER2-FcC-EGFR in 6hr 0.5mM IPTG induced fraction; lane 4, Total 30℃ HER2-FcC-EGFR overnight uninduced fraction; lane 5, Total 30℃ HER2-FcC-EGFR overnight 0.5mM IPTG induced fraction; lane 6, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 7, Total 16℃ HER2-FcC-EGFR overnight uninduced fraction; lane 8, Total 16℃ HER2-FcC-EGFR overnight 0.5mM IPTG induced fraction. ON = overnight. | ||
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| + | :::https://static.igem.wiki/teams/5271/part-figures/sds-gel-her2-fc-egfr-25c.png | ||
| + | :::'''Figure 3. Protein expression of HER2-FC-EGFRnb.''' Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Total 25℃ HER2-FC-EGFR-1 in 6hr uninduced fraction; lane 3, Total 25℃ HER2-FC-EGFR-1 in 6hr 0.5mM IPTG induced fraction; lane 4, Total 25℃ HER2-FC-EGFR-1 overnight uninduced fraction; lane 5, Total 25℃ HER2-FC-EGFR-1 overnight 0.5mM IPTG induced fraction; lane 6, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 7, Total 25℃ HER2-FcC-EGFR-2 in 6hr uninduced fraction; lane 8, Total 25℃ HER2-FcC-EGFR-2 in 6hr 0.5mM IPTG induced fraction; lane 9, Total 25℃ HER2-FcC-EGFR-2 overnight uninduced fraction; lane 10, Total 25℃ HER2-FcC-EGFR-2 overnight 0.5mM IPTG induced fraction. ON = overnight, hr= Hours | ||
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| + | :::https://static.igem.wiki/teams/5271/part-figures/sds-gel-her2-fc-egfr-25c-sol-insol.png | ||
| + | :::'''Figure 4. Protein expression of HER2-FcC-EGFRnb.''' Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Soluble 25℃ HER2-FcC-EGFR-1 in 6hr 0.5mM IPTG induced soluble fraction; lane 3, Insoluble 25℃ HER2-FcC-EGFR-1 in 6hr IPTG induced insoluble fraction; lane 4, Soluble 25℃ HER2-FcC-EGFR-1 overnight IPTG induced soluble fraction; lane 5, Insoluble 25℃ HER2-FcC-EGFR-1 overnight IPTG induced insoluble fraction; lane 6, Soluble 25℃ HER2-FcC-EGFR-2 in 6hr IPTG induced soluble fraction; lane 7, Insoluble 25℃ HER2-FcC-EGFR-2 in 6hr IPTG induced insoluble fraction; lane 8, Soluble 25℃ HER2-FcC-EGFR-2 overnight IPTG induced soluble fraction; lane 9, Insoluble 25℃ HER2-FcC-EGFR-2 overnight IPTG induced insoluble fraction; lane 10, Soluble 25℃ HER2-FcC-EGFR-2 in 6hr uninduced soluble fraction. ON = overnight; Sol = soluble; Insol = insoluble | ||
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| + | :::https://static.igem.wiki/teams/5271/part-figures/sds-gel-her2-fc-egfr-16c-25c-sol-insol.png | ||
| + | :::'''Figure 5. Protein expression of HER2-FcC-EGFR. Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Soluble HER2-FcC-EGFR in 16℃ with 0.2mM IPTG induced fraction; lane 3, Soluble HER2-FcC-EGFR in 16℃ with uninduced fraction; lane 4, Soluble HER2-FcC-EGFR in 16℃ with 0.5mM IPTG induced fraction; lane 5, Soluble HER2-FcC-EGFR in 16℃ with uninduced fraction; lane 6, Soluble HER2-FcC-EGFR in 25℃ with 0.2mM IPTG induced fraction; lane 7, Soluble HER2-FcC-EGFR in 25℃ uninduced fraction; lane 8, Insoluble HER2-FcC-EGFR in 16℃ with 0.2mM uninduced fraction; lane 9, Insoluble HER2-FcC-EGFR in 16℃ with 0.5mM IPTG induced fraction; lane 10, Insoluble HER2-FcC-EGFR e in 25℃ with 0.2mM uninduced fraction. Sol=soluble; Insol = insoluble | ||
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| + | =Reference= | ||
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Revision as of 09:41, 30 September 2024
HER2nb-Fc-EGFRnb
A nanobody composed of two binding regions of EGFR and HER2 linked by a fragment of Fc region of the heavy chain of human IgG.
Profile
- Name: HER2nb-Fc-EGFRnb
- Base Pairs: 1515 bp
- Amino acid: 527 a.a
- Origin: Synthetic
- Property: A dual targeting nanobody with a molecular weight of 55 kDa consisting of a Fc fragment and two HER2 and EGFR nanobody binding domains that specifically bind to EGFR and HER2.
Usage and Biology
Expression
- Figure 2. Protein expression of HER2-FcC-EGFR. Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Total 30℃ HER2-FcC-EGFR in 6hr uninduced fraction; lane 3, Total 30℃ HER2-FcC-EGFR in 6hr 0.5mM IPTG induced fraction; lane 4, Total 30℃ HER2-FcC-EGFR overnight uninduced fraction; lane 5, Total 30℃ HER2-FcC-EGFR overnight 0.5mM IPTG induced fraction; lane 6, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 7, Total 16℃ HER2-FcC-EGFR overnight uninduced fraction; lane 8, Total 16℃ HER2-FcC-EGFR overnight 0.5mM IPTG induced fraction. ON = overnight.
- Figure 3. Protein expression of HER2-FC-EGFRnb. Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Total 25℃ HER2-FC-EGFR-1 in 6hr uninduced fraction; lane 3, Total 25℃ HER2-FC-EGFR-1 in 6hr 0.5mM IPTG induced fraction; lane 4, Total 25℃ HER2-FC-EGFR-1 overnight uninduced fraction; lane 5, Total 25℃ HER2-FC-EGFR-1 overnight 0.5mM IPTG induced fraction; lane 6, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 7, Total 25℃ HER2-FcC-EGFR-2 in 6hr uninduced fraction; lane 8, Total 25℃ HER2-FcC-EGFR-2 in 6hr 0.5mM IPTG induced fraction; lane 9, Total 25℃ HER2-FcC-EGFR-2 overnight uninduced fraction; lane 10, Total 25℃ HER2-FcC-EGFR-2 overnight 0.5mM IPTG induced fraction. ON = overnight, hr= Hours
- Figure 4. Protein expression of HER2-FcC-EGFRnb. Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Soluble 25℃ HER2-FcC-EGFR-1 in 6hr 0.5mM IPTG induced soluble fraction; lane 3, Insoluble 25℃ HER2-FcC-EGFR-1 in 6hr IPTG induced insoluble fraction; lane 4, Soluble 25℃ HER2-FcC-EGFR-1 overnight IPTG induced soluble fraction; lane 5, Insoluble 25℃ HER2-FcC-EGFR-1 overnight IPTG induced insoluble fraction; lane 6, Soluble 25℃ HER2-FcC-EGFR-2 in 6hr IPTG induced soluble fraction; lane 7, Insoluble 25℃ HER2-FcC-EGFR-2 in 6hr IPTG induced insoluble fraction; lane 8, Soluble 25℃ HER2-FcC-EGFR-2 overnight IPTG induced soluble fraction; lane 9, Insoluble 25℃ HER2-FcC-EGFR-2 overnight IPTG induced insoluble fraction; lane 10, Soluble 25℃ HER2-FcC-EGFR-2 in 6hr uninduced soluble fraction. ON = overnight; Sol = soluble; Insol = insoluble
- Figure 5. Protein expression of HER2-FcC-EGFR. Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Soluble HER2-FcC-EGFR in 16℃ with 0.2mM IPTG induced fraction; lane 3, Soluble HER2-FcC-EGFR in 16℃ with uninduced fraction; lane 4, Soluble HER2-FcC-EGFR in 16℃ with 0.5mM IPTG induced fraction; lane 5, Soluble HER2-FcC-EGFR in 16℃ with uninduced fraction; lane 6, Soluble HER2-FcC-EGFR in 25℃ with 0.2mM IPTG induced fraction; lane 7, Soluble HER2-FcC-EGFR in 25℃ uninduced fraction; lane 8, Insoluble HER2-FcC-EGFR in 16℃ with 0.2mM uninduced fraction; lane 9, Insoluble HER2-FcC-EGFR in 16℃ with 0.5mM IPTG induced fraction; lane 10, Insoluble HER2-FcC-EGFR e in 25℃ with 0.2mM uninduced fraction. Sol=soluble; Insol = insoluble
Reference
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 154
Illegal NheI site found at 1615 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 643
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 820
Illegal AgeI site found at 1275
Illegal AgeI site found at 1407 - 1000COMPATIBLE WITH RFC[1000]