Difference between revisions of "Part:BBa K2275010"

Revision as of 15:24, 27 October 2017

PIacI-B0034-K2275004 (glnA-gudB)

We combined the gudB and glnA gene into pSB1C3 backbone. By SDS-PAGE method, we test the expression of both proteins as well. The gel image shows below, where "WC" indicates whole cell, "S" for supernatant, "P" for pellet, and "C" indicates the control of sample without plasmid.

We can see that the glnA and gudB both expressed in E. coli at the same time. Next, we want to test if the Biobrick can function to convert ammonium to glutamine, so we add different ammonium concentration and use kit to detect the glutamine concentration.

The figure above is the functional test result. When merely adding ammonium, the concentration of glutamine is very low. However, if both harboring the plasmid and adding ammonium, the concentration if glutamine is higher.The second and third bar can show the ability of converting ammonium to glutamine. Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 675
Illegal BamHI site found at 1309
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 322
Illegal NgoMIV site found at 1088
Illegal NgoMIV site found at 2454
Illegal AgeI site found at 578
Illegal AgeI site found at 2891
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 1481

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2275010 short</partinfo>
-glnA+rev gudB
+We combined the gudB and glnA gene into pSB1C3 backbone. By SDS-PAGE method, we test the expression of both proteins as well. The gel image shows below, where "WC" indicates whole cell, "S" for supernatant, "P" for pellet, and "C" indicates the control of sample without plasmid.
+[[File:SDS-PAGE_result_of_GlnA_and_GudB.png|220px||centre]]
+We can see that the glnA and gudB both expressed in E. coli at the same time. Next, we want to test if the Biobrick can function to convert ammonium to glutamine, so we add different ammonium concentration and use kit to detect the glutamine concentration.
+[[File:Functional_test_of_glnA-gudB.png|220px||centre]]
+The figure above is the functional test result. When merely adding ammonium, the concentration of glutamine is very low. However, if both harboring the plasmid and adding ammonium, the concentration if glutamine is higher.The second and third bar can show the ability of converting ammonium to glutamine.
 <!-- Add more about the biology of this part here
 ===Usage and Biology===