Difference between revisions of "Part:BBa K3504009"

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==Characterization==
 
==Characterization==
 
==Improvements==
 
==Improvements==
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Using information in literature we were able to increase the replicon cloning and functional ability by adding Q739L Mutation to NSP2
+
Using information in literature we were able to increase the replicon cloning and functional ability by adding G110G, G763R Mutation to NSP2 and adding K94E, S243G,E255D,V305M Mutation to NSP3
−
[[Image:Q739L_Mutation.png|thumb|right|Figure 1. Q739L Mutation in Nsp2.]]
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 +
[[Image:Nsp2-G110G.PNG|thumb|left|Figure 1. Nsp2 G110G Amino acid mutation.]]
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[[Image:Nsp2-G763R.PNG|thumb|Right|Figure 2. Nsp2 G763R Amino acid mutation.]]
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[[Image:Nsp3-K94E.PNG|thumb|left|Figure 3. Nsp3 K94E Amino acid mutation.]]
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[[Image:Nsp3-S243G.PNG|thumb|right|Figure 4. Nsp3 S243G Amino acid mutation.]]
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[[Image:Nsp3-E255D.PNG|thumb|left|Figure 5. Nsp3 E255D Amino acid mutation.]]
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[[Image:Nsp3-V305M.PNG|thumb|right|Figure 6. Nsp3 V305M Amino acid mutation.]]
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==References==
 +
 
 +
Li, Y., Teague, B., Zhang, Y., Su, Z., Porter, E., Dobosh, B., ... & Weiss, R. (2019). In vitro evolution of enhanced RNA replicons for immunotherapy. Scientific reports, 9(1), 1-10.
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 08:05, 13 October 2020


Alphavirus replicon NSPs- Equine Encephalitis virus

Part Description

A composite of parts (BBa_K3504000,BBa_K3504001,BBa_K3504002,BBa_K3504003) and CMV promoter which form ,as a unit, the main constituent of alphavirus replicon which as a whole can give the circuit self-replicating ability

Usage

Characterization

Improvements

Using information in literature we were able to increase the replicon cloning and functional ability by adding G110G, G763R Mutation to NSP2 and adding K94E, S243G,E255D,V305M Mutation to NSP3

Figure 1. Nsp2 G110G Amino acid mutation.
Figure 2. Nsp2 G763R Amino acid mutation.
Figure 3. Nsp3 K94E Amino acid mutation.
Figure 4. Nsp3 S243G Amino acid mutation.
Figure 5. Nsp3 E255D Amino acid mutation.
Figure 6. Nsp3 V305M Amino acid mutation.

References

Li, Y., Teague, B., Zhang, Y., Su, Z., Porter, E., Dobosh, B., ... & Weiss, R. (2019). In vitro evolution of enhanced RNA replicons for immunotherapy. Scientific reports, 9(1), 1-10. Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2760
    Illegal SpeI site found at 2708
    Illegal PstI site found at 5089
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2760
    Illegal NheI site found at 5960
    Illegal NheI site found at 7290
    Illegal NheI site found at 8107
    Illegal SpeI site found at 2708
    Illegal PstI site found at 5089
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2760
    Illegal BglII site found at 2882
    Illegal BamHI site found at 2744
    Illegal BamHI site found at 3107
    Illegal XhoI site found at 6124
    Illegal XhoI site found at 6185
    Illegal XhoI site found at 6226
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2760
    Illegal SpeI site found at 2708
    Illegal PstI site found at 5089
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2760
    Illegal SpeI site found at 2708
    Illegal PstI site found at 5089
    Illegal NgoMIV site found at 3549
    Illegal AgeI site found at 5665
    Illegal AgeI site found at 6034
    Illegal AgeI site found at 6361
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1635
    Illegal BsaI site found at 6223
    Illegal BsaI site found at 6241
    Illegal BsaI.rc site found at 3012
    Illegal BsaI.rc site found at 3659
    Illegal BsaI.rc site found at 5750