Difference between revisions of "Part:BBa K4115021"

 
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K4115021 short</partinfo>
 
<partinfo>BBa_K4115021 short</partinfo>
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{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
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! colspan="2" style="background:#FFBF00;"|glgA gene silence
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|-
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|'''Function'''
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|Gene silence
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|-
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|'''Use in'''
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|<i>S.elongatus</i> HL7942<br>
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|-
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|'''RFC standard'''
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|None<br>
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|-
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|'''Backbone'''
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|pUC57<br>
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|-
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|'''Submitted by'''
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|[https://2022.igem.wiki/shanghaitech-china/ ShanghaiTech_China]
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|}
  
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This Composite part is integrated into the plasmid backbone of PUC57 and entered Synechococcus by natural transformation with E. coli. The glgA sRNA in the part will transcribe small RNA fragments that can bind specifically to glgA's mRNA to inhibit the expression of glgA. The silence of glgA will inhibit G1P's formation into glycogen and promotes its formation into sucrose, which increases the synthesis of Synechococcus sucrose. This allows Synechococcus to transport more sucrose via cscB in response to E. coli carbon starvation signals.
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This composite part entered <i>S.elongatus</i> HL7942 by natural transformation with <i>E. coli</i> with the help of two homology arms NSI-US [https://parts.igem.org/Part:BBa_K4115010 (BBa_K4115010)] and NSI-DS [https://parts.igem.org/Part:BBa_K4115011 (BBa_K3971011)]. The glgA sRNA [https://parts.igem.org/Part:BBa_K4115015 (BBa_K4115015)] in the part will transcribe small RNA fragments that can bind specifically to glgA's mRNA to inhibit the expression of glgA assisted by HFQ [https://parts.igem.org/Part:BBa_K1963000 (BBa_K1963000)] [1].<br>
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The silence of glgA will inhibit G1P's formation into glycogen and promotes its formation into sucrose, which increases the synthesis of <i>S.elongatus</i> HL7942 sucrose [2]. This allows HL7942 to transport more sucrose via cscB in response to <i>E. coli</i> carbon starvation signals.
  
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
  
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<!-- -->
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When no starvation signal is received, LacI is expressed and binds to the lac operator on the lac UV5 promoter [https://parts.igem.org/Part:BBa_K4115012 (BBa_K4115012)] to inhibit the expression of T7 RNA polymerase [https://parts.igem.org/Part:BBa_K145001 (BBa_K145001)], thus glgA sRNA was not synthesized. When a starvation signal is received, the amount of LacI decreases, so that the lac UV5 promoter is activated to express the downstream T7 RNA polymerase. With T7 RNA polymerase, the T7 promoter [https://parts.igem.org/Part:BBa_I719005 (BBa_I719005)] is activated to express their respective downstream HFQ [https://parts.igem.org/Part:BBa_K1963000 (BBa_K1963000)] and glgA sRNA [https://parts.igem.org/Part:BBa_K4115015 (BBa_K4115015)]. With the help of the HFQ protein, the sRNA will specifically bind to the mRNA of glgA, so that it inhibits the expression of the glgA gene.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K4115021 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4115021 SequenceAndFeatures</partinfo>
  
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== References ==
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<small>
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[1] Li, S., Sun, T., Chen, L., Zhang, W. (2021). Designing and Constructing Artificial Small RNAs for Gene Regulation and Carbon Flux Redirection in Photosynthetic Cyanobacteria. In: Basu, C. (eds) Biofuels and Biodiesel. Methods in Molecular Biology, vol 2290. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1323-8_16
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[2] Lin PC, Zhang F, Pakrasi HB. Enhanced production of sucrose in the fast-growing cyanobacterium Synechococcus elongatus UTEX 2973. Sci Rep. 2020 Jan 15;10(1):390. doi: 10.1038/s41598-019-57319-5. PMID: 31942010; PMCID: PMC6962321.
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 10:42, 4 October 2022


glgA gene silence

glgA gene silence
Function Gene silence
Use in S.elongatus HL7942
RFC standard None
Backbone pUC57
Submitted by ShanghaiTech_China

This composite part entered S.elongatus HL7942 by natural transformation with E. coli with the help of two homology arms NSI-US (BBa_K4115010) and NSI-DS (BBa_K3971011). The glgA sRNA (BBa_K4115015) in the part will transcribe small RNA fragments that can bind specifically to glgA's mRNA to inhibit the expression of glgA assisted by HFQ (BBa_K1963000) [1].
The silence of glgA will inhibit G1P's formation into glycogen and promotes its formation into sucrose, which increases the synthesis of S.elongatus HL7942 sucrose [2]. This allows HL7942 to transport more sucrose via cscB in response to E. coli carbon starvation signals.


Usage and Biology

When no starvation signal is received, LacI is expressed and binds to the lac operator on the lac UV5 promoter (BBa_K4115012) to inhibit the expression of T7 RNA polymerase (BBa_K145001), thus glgA sRNA was not synthesized. When a starvation signal is received, the amount of LacI decreases, so that the lac UV5 promoter is activated to express the downstream T7 RNA polymerase. With T7 RNA polymerase, the T7 promoter (BBa_I719005) is activated to express their respective downstream HFQ (BBa_K1963000) and glgA sRNA (BBa_K4115015). With the help of the HFQ protein, the sRNA will specifically bind to the mRNA of glgA, so that it inhibits the expression of the glgA gene.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 333
    Illegal XbaI site found at 1903
    Illegal PstI site found at 6912
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 520
    Illegal PstI site found at 6912
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 333
    Illegal XbaI site found at 1903
    Illegal PstI site found at 6912
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 333
    Illegal XbaI site found at 1903
    Illegal PstI site found at 6912
    Illegal AgeI site found at 2068
    Illegal AgeI site found at 6284
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2065
    Illegal BsaI.rc site found at 6281

References

[1] Li, S., Sun, T., Chen, L., Zhang, W. (2021). Designing and Constructing Artificial Small RNAs for Gene Regulation and Carbon Flux Redirection in Photosynthetic Cyanobacteria. In: Basu, C. (eds) Biofuels and Biodiesel. Methods in Molecular Biology, vol 2290. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1323-8_16

[2] Lin PC, Zhang F, Pakrasi HB. Enhanced production of sucrose in the fast-growing cyanobacterium Synechococcus elongatus UTEX 2973. Sci Rep. 2020 Jan 15;10(1):390. doi: 10.1038/s41598-019-57319-5. PMID: 31942010; PMCID: PMC6962321.