Plasmid

Part:BBa_K5302017

Designed by: Xinyuan Shi   Group: iGEM24_USTC   (2024-10-01)
Revision as of 09:54, 1 October 2024 by Xiaotian (Talk | contribs)


pBBR-OmpA-8IIU

8IIU is an anti-VEGF nanobody, and it has 2 chains, chainA sequence: mdvqlvesgg glvqpggslr lscaasgrtf ssysmgwfrq apgkerefvv aiskggykyd avslegrfti srdnakntvy lqinslrped tavyycassr aygssrlrla dtyeywgqgt lvtvss. chainB sequence:mdvqlvesgg glvqpggslr lscaasgrtf ssysmgwfrq apgkerefvv aiskggyky avslegrfti srdnakntvy lqinslrped tavyycassr aygssrlrla dtyeywgqgt lvtvss.

These two chains all show great affinity with VEGF, we used pBBR plasmid as a backbone and transfered 8IIU into Escherichia coli Nissle 1917, and finally succeeded in expressing 8IIU.

Jamboree Program
Figure 1. Colony PCR results of pBBR-OmpA-8IIU

Jamboree Program
Figure 2. SDS-PAGE analysis of pBBR1MCS-INP-miniZ expression in Escherichia coli Nissle 1917

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3931
    Illegal EcoRI site found at 4301
    Illegal XbaI site found at 3140
    Illegal PstI site found at 1967
    Illegal PstI site found at 3128
    Illegal PstI site found at 4033
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3931
    Illegal EcoRI site found at 4301
    Illegal PstI site found at 1967
    Illegal PstI site found at 3128
    Illegal PstI site found at 4033
    Illegal NotI site found at 1008
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3931
    Illegal EcoRI site found at 4301
    Illegal BglII site found at 1754
    Illegal BamHI site found at 3146
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 3931
    Illegal EcoRI site found at 4301
    Illegal XbaI site found at 3140
    Illegal PstI site found at 1967
    Illegal PstI site found at 3128
    Illegal PstI site found at 4033
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 3931
    Illegal EcoRI site found at 4301
    Illegal XbaI site found at 3140
    Illegal PstI site found at 1967
    Illegal PstI site found at 3128
    Illegal PstI site found at 4033
    Illegal NgoMIV site found at 2418
    Illegal NgoMIV site found at 2701
    Illegal NgoMIV site found at 4959
    Illegal AgeI site found at 4799
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1330
    Illegal SapI.rc site found at 2267
    Illegal SapI.rc site found at 2477


[edit]
Categories
//plasmid
Parameters
biologyEscherichia coli Nissle 1917