RBS

Part:BBa_B0033

Designed by: Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.   Group: Antiquity   (2003-01-31)
Revision as of 15:22, 20 October 2019 by Ezio1999 (Talk | contribs)

RBS.4 (weaker) -- derivative of BBa_0030

Weaker RBS based on Ron Weiss thesis. Strengths relative to BBa_B0030, BBa_B0031, BBa_B0032.

Improvement

Group:iGEM2019_JiangnanU_China

This year, we improved the sequence of the RBS BBa_B0033, and got the RBS BBa_K3137001.

JiangnanUchina improve 1.png

As we all know, RBS refers to a sputum-rich untranslated region upstream of the initiation codon AUG. BBa_B0033 is a weaker RBS based on Ron Weiss thesis. Compared with BBa_B0030, BBa_B0031, BBa_B0032, BBa_B0034, BBa_B0033 is weakest, and whose RBS strength is 0.35% of BBa_B0034. By analyzing their sequence, we found that BBa_B0033 has fewer purines and no not have AAA sequence. SO we designed the RBS by adjusting the proportion of bases.

JiangnanUchina improve 2.png

Then we ligated the two RBS with promotor rsmH (BBa_K3137000) and gene gfp (BBa_K3137011) and transform into E. coli BL21 to compare their green fluorescence. And take E. coli BL21 as a control. The figure below shows that BBa_K3137001 has the stronger fluorescently protein expression than BBa_B0033, that means it’s efficient to increase the protein expression by increasing the number of purines in the RBS sequence, which is beneficial to the binding of ribosomes. JiangnanUchina improve 3.png JiangnanUchina improve 4.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
    COMPATIBLE WITH RFC[1000]

Functional Parameters

biology-NA-
efficiency0.01

(Relative to BBa_B0034)

Team Warsaw 2010's measurement

RBS strength (relative to B0034): 0,35%

[edit]
Categories
//rbs/prokaryote/constitutive/community
//ribosome/prokaryote/ecoli
//chassis/prokaryote/ecoli
//direction/forward
//regulation/constitutive
Parameters
biology
efficiency0.01