Composite

Part:BBa_K5271012

Designed by: Yin Yan Chan   Group: iGEM24_HKPOLYU   (2024-09-30)
Revision as of 09:59, 30 September 2024 by Carriechan1 (Talk | contribs)


sec-HER2nb-Fc-EGFRnb

A nanobody of HERnb-Fc-EGFRnb carrying a secretory peptide for improved secretion



Profile

  • Name:sec-HER2nb-Fc-EGFRnb
  • Base Pairs: 1581 bp
  • Amino acid: 527 a.a
  • Origin: Synthetic
  • Property:



Usage and Biology

sec-her2-fc-egfr-structure1.png
Figure The AlphaFold predicted structure of sec-HER2-Fc-EGFR. Cyan: signal peptide ssSTII1 S-13L; Red: His-tag; Green: HER2 nanobody; Grey: linker; Blue: Fc protein; Orange: EGFR nanobody.




Expression

sds-gel-sec-her2-fc-egfr-16c-25c-30c-total.png
Figure 2. Protein expression of sec-HER2-Fc-EGFR. Lane 1, Total sec-HER2-Fc-EGFR in 16℃ with 0.5mM IPTG induced fraction for 24 hours; lane 2, Total sec-HER2-Fc-EGFR in 25℃ with 0.5m IPTG induced fraction for 24 hours; lane 3, Total sec-HER2-Fc-EGFR in 30℃ with 0.5mM IPTG induced fraction for 24 hours; lane 4, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 5, Total sec-HER2-Fc-EGFR in 16℃ with uninduced fraction for 24 hours; lane 6, Total sec-HER2-Fc-EGFR in 25℃ with uninduced fraction for 24 hours; lane 7, Total sec-HER2-Fc-EGFR in 30℃ with uninduced fraction for 24 hours.​



sds-gel-sec-her2-fc-egfr-16c-25c-30c-sol.png
Figure 3. Protein expression of sec-HER2-Fc-EGFR. Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Soluble sec-HER2-Fc-EGFR in 16℃ with 0.5mM IPTG induced fraction for 24 hours; lane 3, Soluble sec-HER2-Fc-EGFR in 25℃ with 0.5m IPTG induced fraction for 24 hours; lane 3, Soluble sec-HER2-Fc-EGFR 5 in 30℃ with 0.5mM IPTG induced fraction for 24 hours; lane 5, Soluble sec-HER2-Fc-EGFR 5 in 16℃ with uninduced fraction for 24 hours; lane 6, Soluble sec-HER2-Fc-EGFR in 25℃ with uninduced fraction for 24 hours; lane 7, Soluble sec-HER2-Fc-EGFR in 30℃ with uninduced fraction for 24 hours.​



sds-gel-sec-her2-fc-egfr-16c-25c-30c-insol.png
Figure 4. Protein expression of sec-HER2-Fc-EGFR. Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Insoluble sec-HER2-Fc-EGFR in 16℃ with 0.5mM IPTG induced fraction for 24 hours; lane 3, Insoluble sec-HER2-Fc-EGFR in 25℃ with 0.5m IPTG induced fraction for 24 hours; lane 3, Insoluble sec-HER2-Fc-EGFR 5 in 30℃ with 0.5mM IPTG induced fraction for 24 hours; lane 5, Insoluble sec-HER2-Fc-EGFR 5 in 16℃ with uninduced fraction for 24 hours; lane 6, Insoluble sec-HER2-Fc-EGFR in 25℃ with uninduced fraction for 24 hours; lane 7, Insoluble sec-HER2-Fc-EGFR in 30℃ with uninduced fraction for 24 hours.​



sds-gel-her2-fc-egfr-4.png
Figure 5. Protein expression of HER2-FcC-EGFR. Lane 1, Biorad Precision Plus Protein™™ Unstained Protein Standards; lane 2, Total HER2-FcC-EGFR in 16℃ with 0.2mM IPTG induced fraction; lane 3, Total HER2-FcC-EGFR in 16℃ with uninduced fraction; lane 4, Total HER2-FcC-EGFR in 16℃ with 0.5mM IPTG induced fraction; lane 5, Total HER2-FcC-EGFR in 16℃ with uninduced fraction; lane 6, Total HER2-FcC-EGFR in 25℃ with 0.2mM IPTG induced fraction; lane 7, Total HER2-FcC-EGFR in 25℃ uninduced fraction; lane 8, Insoluble HER2-FcC-EGFR in 16℃ with 0.2mM IPTG induced fraction; lane 9, Insoluble HER2-FcC-EGFR in 16℃ with uninduced fraction; lane 10, Insoluble HER2-FcC-EGFR in 16℃ with 0.5mM IPTG induced fraction. Insol = insoluble​







Reference

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 228
    Illegal NheI site found at 1689
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 717
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 894
    Illegal AgeI site found at 1349
    Illegal AgeI site found at 1481
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//awards/composite_part
//proteindomain/binding
Parameters
None