Difference between revisions of "Part:BBa K4115021"

Revision as of 08:38, 4 October 2022

glgA gene silence

NS3-2-lac UV5 promoter-cscB-lacI-KanR-NS3-1
Function	Permease
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). 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. 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

@@ Line 2: / Line 2: @@
 __NOTOC__
 <partinfo>BBa_K4115021 short</partinfo>
+{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
+! colspan="2" style="background:#FFBF00;"|NS3-2-lac UV5 promoter-cscB-lacI-KanR-NS3-1
+|-
+|'''Function'''
+|Permease
+|-
+|'''Use in'''
+|<i>S.elongatus</i> HL7942<br>
+|-
+|'''RFC standard'''
+|None<br>
+|-
+|'''Backbone'''
+|pUC57<br>
+|-
+|'''Submitted by'''
+|[https://2022.igem.wiki/shanghaitech-china/ ShanghaiTech_China]
+|}
-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)]. 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.
+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)]. 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. This allows HL7942 to transport more sucrose via cscB in response to <i>E. coli</i> carbon starvation signals.
@@ Line 10: / Line 28: @@
 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.
-<!-- -->
 <span class='h3bb'>Sequence and Features</span>
 <partinfo>BBa_K4115021 SequenceAndFeatures</partinfo>