Difference between revisions of "Part:BBa K3137001"

Revision as of 15:24, 20 October 2019

Improved RBS 4

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

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.

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.

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]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3137001 short</partinfo>
-DnaE is the alpha subunit of DNA polymerase III which catalyzes the polymerase activity of the holoenzyme complex.
+This year, we improved the sequence of the RBS <bbpart>BBa_B0033</bbpart>, and got the RBS <bbpart>BBa_K3137001</bbpart>.
+[[Image:JiangnanUchina improve 1.png|400px|center]]
+As we all know, RBS refers to a sputum-rich untranslated region upstream of the initiation codon AUG. <bbpart>BBa_B0033</bbpart> is a weaker RBS based on Ron Weiss thesis. Compared with <bbpart>BBa_B0030</bbpart>, <bbpart>BBa_B0031</bbpart>, <bbpart>BBa_B0032</bbpart>, <bbpart>BBa_B0034</bbpart>, <bbpart>BBa_B0033</bbpart> is weakest, and whose RBS strength is 0.35% of <bbpart>BBa_B0034</bbpart>. By analyzing their sequence, we found that <bbpart>BBa_B0033</bbpart> has fewer purines and no not have AAA sequence. SO we designed the RBS by adjusting the proportion of bases.
+[[Image:JiangnanUchina improve 2.png|400px|center]]
+Then we ligated the two RBS with promotor rsmH (<bbpart>BBa_K3137000</bbpart>) and gene gfp (<bbpart>BBa_K3137011</bbpart>) 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.
+[[Image:JiangnanUchina improve 3.png|400px|]]
+[[Image:JiangnanUchina improve 4.png|400px|]]
 <!-- Add more about the biology of this part here