Part:BBa_K4115021
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
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 333
Illegal XbaI site found at 1903
Illegal PstI site found at 6912 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 520
Illegal PstI site found at 6912 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 333
Illegal XbaI site found at 1903
Illegal PstI site found at 6912 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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.
| None |