Difference between revisions of "Part:BBa K2970008"

Revision as of 12:02, 20 October 2019

Gate/Trigger test composition

To measure the functionality of our gate (BBa_K2970002) and trigger (BBa_K2970000 and BBa_K2970001) we designed a test plasmid which contains both trigger and the gate, which is locking GFP, each embedded into a promoter and a terminator. The promoters of the trigger can be induced by either IPTG or arabinose to test different combinations of trigger concentrations. In contrast, the gate is translated constitutively.

The function of the gate is the regulation of the translation of a gene of interest, in this case GFP for testing, by formation of a hairpin by the mRNA that hides the ribosome binding site and the start codon. The mRNAs of the two triggers can form a complex due to complementarity, which can open the gates hairpin and releases the ribosome binding site, thus translation can take place. The affinity of the trigger complex is higher than the affinity of the gate within. The gate can only be opened if both triggers are present, otherwise the complementary region is too short. If the gate is opened, GFP is expressed whose signal can be measured.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 103
Illegal NheI site found at 284
Illegal NheI site found at 422
Illegal NheI site found at 445
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 224
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1219

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2970008 short</partinfo>
-It contains both triggers and the gate to test all three. (<partinfo>BBa_K2970000</partinfo>, <partinfo>BBa_K2970001</partinfo>, <partinfo>BBa_K2970002</partinfo>). The function of the gate is the regulation of the translation, which is activated by the triggers.
+To measure the functionality of our gate (<partinfo>BBa_K2970002</partinfo>) and trigger (<partinfo>BBa_K2970000</partinfo> and <partinfo>BBa_K2970001</partinfo>) we designed a test plasmid which contains both trigger and the gate, which is locking GFP, each embedded into a promoter and a terminator. The promoters of the trigger can be induced by either IPTG or arabinose to test different combinations of trigger concentrations. In contrast, the gate is translated constitutively.
-The two triggers form a complex due to complementary areas. The sequence that is formed is complementary to the gate that in normal conformation forms a hairpin which covers the RBS and the start codon. If the trigger complex binds to the gate, this hairpin opens and the RBS is released and translation can take place. The affinity of the trigger complex is higher than the affinity of the gate within. The gate can only be opened if both triggers are present, otherwise the complementary region is too short. If the gate is opened, GFP is expressed whose signal can be measured.
+The function of the gate is the regulation of the translation of a gene of interest, in this case GFP for testing, by formation of a hairpin by the mRNA that hides the ribosome binding site and the start codon. The mRNAs of the two triggers can form a complex due to complementarity, which can open the gates hairpin and releases the ribosome binding site, thus translation can take place. The affinity of the trigger complex is higher than the affinity of the gate within. The gate can only be opened if both triggers are present, otherwise the complementary region is too short. If the gate is opened, GFP is expressed whose signal can be measured.
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