Difference between revisions of "Part:BBa K2213006"
Line 17: Line 17:
 
<partinfo>BBa_K2213006 parameters</partinfo>
 
<partinfo>BBa_K2213006 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
Function:
 +
<br>
 +
This part is an improved version of BBa_K562001, originally submitted by team Dundee 2011. The original part contained an illegal XbaI site which has been removed to make it biobrick compatible. This part has been expressed under different strength promoters. A low strength Anderson promoter here, medium strength Anderson promoter (BBa_K2213007), and a high strength Anderson promoter (BBa_K2213008).<br>
 +
<br>
 +
The PduD tag was combined with the low strength Anderson promoter (BBa_J23105) and mCherry.<br>
 +
<br>
 +
https://static.igem.org/mediawiki/2017/e/e4/PromoterComparison800p.png<br>
 +
<br>
 +
A gradient of fluorescence is evident when compared to medium and high promoters.<br>
 +
<br>
 +
<br>
 +
<br>
 +
https://static.igem.org/mediawiki/2017/b/bd/TagExpression800p.jpeg
 +
<br>
 +
The expression levels compared to medium and high are as shown.<br>
 +
<br>
 +
<br>
 +
<br>
 +
https://static.igem.org/mediawiki/2017/1/16/LowPromoterComparison800p.png<br>
 +
<br>
 +
When expressed alone the distribution of the mCherry signal was homogeneous and fluorescence level was very low making visualisation difficult. In the presence of EutSMNLK the mCherry signal was much brighter and clumped together, indicating localisation to the BMC.

Revision as of 20:31, 30 October 2017


LowPromoter_PduD(1-20)_mCherry

This part is an improved version of https://parts.igem.org/Part:BBa_K562001, submitted by Dundee 2011.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 8
    Illegal NheI site found at 31
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Function:
This part is an improved version of BBa_K562001, originally submitted by team Dundee 2011. The original part contained an illegal XbaI site which has been removed to make it biobrick compatible. This part has been expressed under different strength promoters. A low strength Anderson promoter here, medium strength Anderson promoter (BBa_K2213007), and a high strength Anderson promoter (BBa_K2213008).

The PduD tag was combined with the low strength Anderson promoter (BBa_J23105) and mCherry.

PromoterComparison800p.png

A gradient of fluorescence is evident when compared to medium and high promoters.



TagExpression800p.jpeg
The expression levels compared to medium and high are as shown.



LowPromoterComparison800p.png

When expressed alone the distribution of the mCherry signal was homogeneous and fluorescence level was very low making visualisation difficult. In the presence of EutSMNLK the mCherry signal was much brighter and clumped together, indicating localisation to the BMC.