Difference between revisions of "Part:BBa K2890001"
 
(6 intermediate revisions by the same user not shown)
Line 4: Line 4:
  
 
A optimized frmR promoter for formaldehyde detection.
 
A optimized frmR promoter for formaldehyde detection.
 +
<br/>
 +
<br/>
 +
=Characterization from JNFLS2019=
 +
==1.The toxic effect of formaldehyde on the cells growth==
  
−
 
+
We added the reporter gene GFP to the downstream of the promoter, then detected the tolerance of the cells and the sensitivity of the promoter to formaldehyde using BL21 as chassis.  
−
Contributions from Yuhan Sun, JNFLS 2019.
+
Formaldehyde is toxic to cells. It can inhibit the cells growth if it is added to the culture medium too early. In order to know at which OD600 the formaldehyde should be added into the culture medium, we used 0.5mM formaldehyde to treat cells at different original OD600, then we collected different group bacteria at certain time to measure cells’ DO600 absorbance (Figure 1). The result showed that the toxic effect of formaldehyde on the cells growth was decreased with the cells original OD600 increased.
−
 
+
[[File:K2890001-1.jpg|center]]
−
We added the reporter gene GFP to the downstream of the promoter, then detected the sensitivity and tolerance of the promoter to formaldehyde using BL21 as chassis. We cultured E.coli BL21 transformed with the plasmid （PfrmR-GFP）and added different concentration formaldehyde to different groups when the OD600 = 1, then we collected bacteria at certain time to measure the fluorescence and the absorbance. From the Fig. 1, we found that 0.8 mM formaldehyde is the best concentration for expression reporter protein GFP.
+
==2.The sensitivity of promoter to formaldehyde==
−
 
+
We cultured E.coli BL21 transformed with the plasmid (PfrmR-GFP) and added different concentration formaldehyde to different groups when the OD600 = 1, then we collected bacteria at certain time to measure the fluorescence and the absorbance. From the Figure 2, we found that 0.8 mM formaldehyde is the best concentration for inducing GFP expression.
−
[[File:K2890001_contributions.png|center]]
+
[[File:K2890001-2.jpg|center]]
 +
<br/>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 03:58, 20 October 2019


optimized frmR

A optimized frmR promoter for formaldehyde detection.

Characterization from JNFLS2019

1.The toxic effect of formaldehyde on the cells growth

We added the reporter gene GFP to the downstream of the promoter, then detected the tolerance of the cells and the sensitivity of the promoter to formaldehyde using BL21 as chassis. Formaldehyde is toxic to cells. It can inhibit the cells growth if it is added to the culture medium too early. In order to know at which OD600 the formaldehyde should be added into the culture medium, we used 0.5mM formaldehyde to treat cells at different original OD600, then we collected different group bacteria at certain time to measure cells’ DO600 absorbance (Figure 1). The result showed that the toxic effect of formaldehyde on the cells growth was decreased with the cells original OD600 increased.

K2890001-1.jpg

2.The sensitivity of promoter to formaldehyde

We cultured E.coli BL21 transformed with the plasmid (PfrmR-GFP) and added different concentration formaldehyde to different groups when the OD600 = 1, then we collected bacteria at certain time to measure the fluorescence and the absorbance. From the Figure 2, we found that 0.8 mM formaldehyde is the best concentration for inducing GFP expression.

K2890001-2.jpg


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 SapI.rc site found at 551