Difference between revisions of "Part:BBa K3376014"
 
(5 intermediate revisions by the same user not shown)
Line 5: Line 5:
 
This part was modified from GFPmut1 ([https://parts.igem.org/Part:BBa_K1159311 Part:BBa_K1159311]) designed by TU-Munich in iGEM 2013 by adding ATG and a stop codon for protein expression.  
 
This part was modified from GFPmut1 ([https://parts.igem.org/Part:BBa_K1159311 Part:BBa_K1159311]) designed by TU-Munich in iGEM 2013 by adding ATG and a stop codon for protein expression.  
  
−
=== Research ===
+
=== Expression ===
−
Based on our research, the glucose transporter of ''Salmonella'' has a lower Km compared to human small intestine, ''Staphylococcus'' and ''E. coli'', indicating a higher efficiency for glucose uptake. In our study, we demonstrated glucose absorption ability by overexpressing each of these two systems from ''Salmonella'' in ''E. coli''. Please go to our wiki page ([http://2017.igem.org/Team:Mingdao/Demonstrate#demonstrate-md Mingdao iGEM 2017]) for more information.  
+
GFP was detected at Ex/Em = 483/513. The fluorescence expression levels were measured for lactate dehydrogenase promoter (ldhp) [ldhp-GFP-Tr/pSB1C3 (BBa_K3376002)] and thiol peroxidase promoter (tpxp) [tpxp-GFP-Tr/pSB1C3 (BBa_K3376004)] activity of S. mutans.  
  
−
[[File:Mingdaophil1026-2.png|550px|center]]
+
[[File:T--Mingdao--ww1.png|450px|center]]
  
 +
=== Transformation ===
 +
The ldhp-GFP-Tr/pSB1C3 [BBa_K3376002] was transformed into E. coli Nissle strain. The high expression of green fluorescent protein was observed under a blue led light.
  
−
 
+
[[File:T--Mingdao--ww2.png|600px|center]]
−
=== Cloning ===
+
−
The STM1128 gene was amplified from gDNA of Salmonella typhimurium and cloned onto pSB1C3. This part has been sequenced.
+
−
 
+
−
[[File:Mingdaophil1026-3.jpeg|400px|center]]
+
  
  

Latest revision as of 04:57, 6 October 2020


Aequorea victoria GFPmut1

This part was modified from GFPmut1 (Part:BBa_K1159311) designed by TU-Munich in iGEM 2013 by adding ATG and a stop codon for protein expression.

Expression

GFP was detected at Ex/Em = 483/513. The fluorescence expression levels were measured for lactate dehydrogenase promoter (ldhp) [ldhp-GFP-Tr/pSB1C3 (BBa_K3376002)] and thiol peroxidase promoter (tpxp) [tpxp-GFP-Tr/pSB1C3 (BBa_K3376004)] activity of S. mutans.

T--Mingdao--ww1.png

Transformation

The ldhp-GFP-Tr/pSB1C3 [BBa_K3376002] was transformed into E. coli Nissle strain. The high expression of green fluorescent protein was observed under a blue led light.

T--Mingdao--ww2.png




Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 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