Difference between revisions of "Part:BBa K2243013"

 
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catalyze attL and attR to reset the DNA sequence
 
catalyze attL and attR to reset the DNA sequence
  
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===Usage and Biology===
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===Usage===
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In the last few years, researchers also investigate into integrases work with the help of their Recombination Directionality Factors (RDFs). Bxb1 gp47 is an Bxb1-encoded RDF, the product of gene 47. With gp47, Bxb1 gp35 allows the sequence to be flipped, excised, or inserted between attL and attR sites, which makes it useful for gene editing. We design a fusion protein for the integrase and its RDF. In our project, Bxb1 gp35-gp47 fusion protein can flip the sequence flanked by attL and attR site to reset the sequence flipped by Bxb1 gp35.
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===Biology===
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Integrase Bxb1 gp35 comes from Mycobacteriophage, which allows the phage to insert its DNA into the host’s genome at host’s attP site using viral attB site. With RDF Bxb1 gp47, the recombined attL and attR can be recognized and undergo recombination, which enables the excision of phage genome. We got the sequence by de novo synthesis.
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===Design Notes===
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We assembled the Bxb1 gp35 and gp47 with a linker adapted from a recent research[1] by Gibson Assembly.
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===Source===
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Mycobacterium Phage Bxb1
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===References===
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[1]Olorunniji, F. J., McPherson, A. L., Rosser, S. J., Smith, M. C., Colloms, S. D., & Stark, W. M. (2017). Control of serine integrase recombination directionality by fusion with the directionality factor. Nucleic acids research, 45(14), 8635-8645.
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Revision as of 12:50, 31 October 2017


Bxb1 gp35-gp47 fusion protein

catalyze attL and attR to reset the DNA sequence


Usage

In the last few years, researchers also investigate into integrases work with the help of their Recombination Directionality Factors (RDFs). Bxb1 gp47 is an Bxb1-encoded RDF, the product of gene 47. With gp47, Bxb1 gp35 allows the sequence to be flipped, excised, or inserted between attL and attR sites, which makes it useful for gene editing. We design a fusion protein for the integrase and its RDF. In our project, Bxb1 gp35-gp47 fusion protein can flip the sequence flanked by attL and attR site to reset the sequence flipped by Bxb1 gp35.

Biology

Integrase Bxb1 gp35 comes from Mycobacteriophage, which allows the phage to insert its DNA into the host’s genome at host’s attP site using viral attB site. With RDF Bxb1 gp47, the recombined attL and attR can be recognized and undergo recombination, which enables the excision of phage genome. We got the sequence by de novo synthesis.


Design Notes

We assembled the Bxb1 gp35 and gp47 with a linker adapted from a recent research[1] by Gibson Assembly.


Source

Mycobacterium Phage Bxb1

References

[1]Olorunniji, F. J., McPherson, A. L., Rosser, S. J., Smith, M. C., Colloms, S. D., & Stark, W. M. (2017). Control of serine integrase recombination directionality by fusion with the directionality factor. Nucleic acids research, 45(14), 8635-8645.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 192
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1507
    Illegal BamHI site found at 466
    Illegal XhoI site found at 553
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1105
    Illegal NgoMIV site found at 1192
    Illegal AgeI site found at 242
  • 1000
    COMPATIBLE WITH RFC[1000]