Difference between revisions of "Part:BBa K2047010:Experience"

 
 
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<partinfo>BBa_K2047001 SequenceAndFeatures</partinfo>
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
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how you used this part and how it worked out.
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===Applications of BBa_K2047010===
 
  
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===User Reviews===
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===Design Notes===
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none
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When stem loops inserted in the 3’ end of the upstream gene, it protects its mRNA against the cleavage of exonuclease, increasing the ratio of abundance of the first gene product relative to that of the second gene product. Furthermore, the lower free energy of stem loops are, the more stable the secondary transcripts of the upstream are, tuning the expression of multiple genes. We designed a series of stem loops with different free energy and measured it by fluorescence reporter system.
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===Source===
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<I>Username</I>
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Inspired by Xu’s work and based on keasling’s work, we designed a series of stem loops with different free energy for further use as basic regulatory parts.
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===References===
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<p>[1]. Pfleger, B.F., et al., Combinatorial engineering of intergenic regions in operons tunes expression of multiple genes. Nat Biotechnol, 2006. 24(8): p. 1027-32.<br>[2]. Xu, C., et al., Cellulosome stoichiometry in Clostridium cellulolyticum is regulated by selective RNA processing and stabilization. Nat Commun, 2015. 6: p. 6900.<br>[3]. Smolke, C.D. and J.D. Keasling, Effect of gene location, mRNA secondary structures, and RNase sites on expression of two genes in an engineered operon. Biotechnol Bioeng, 2002. 80(7): p. 762-76.</p>
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Latest revision as of 07:17, 22 October 2016

_NOTOC__ Fluorescent reporter system with stem-loop, GFP_Stem-loop with free energy of -25.6 kcal/mol


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 764
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 644


Design Notes

none When stem loops inserted in the 3’ end of the upstream gene, it protects its mRNA against the cleavage of exonuclease, increasing the ratio of abundance of the first gene product relative to that of the second gene product. Furthermore, the lower free energy of stem loops are, the more stable the secondary transcripts of the upstream are, tuning the expression of multiple genes. We designed a series of stem loops with different free energy and measured it by fluorescence reporter system.

Source

Inspired by Xu’s work and based on keasling’s work, we designed a series of stem loops with different free energy for further use as basic regulatory parts.

References

[1]. Pfleger, B.F., et al., Combinatorial engineering of intergenic regions in operons tunes expression of multiple genes. Nat Biotechnol, 2006. 24(8): p. 1027-32.
[2]. Xu, C., et al., Cellulosome stoichiometry in Clostridium cellulolyticum is regulated by selective RNA processing and stabilization. Nat Commun, 2015. 6: p. 6900.
[3]. Smolke, C.D. and J.D. Keasling, Effect of gene location, mRNA secondary structures, and RNase sites on expression of two genes in an engineered operon. Biotechnol Bioeng, 2002. 80(7): p. 762-76.