Difference between revisions of "Part:BBa K1957002"

Revision as of 21:42, 8 October 2016

HydB (C-terminally tagged) subunit of FeFe Hydrogenase

One of the three subunits that make up FeFe Hydrogenase in Shewanella oneidensis. Specifically this is the HydB subunit which is the central subunit. This basic part has a strep-tag attached to its C-terminus for consequent protein assays. This unit is one out of the three subunits that can be used to make a the entire FeFe hydrogenase construct.

Figure 1: PCR Colony of FeFe Hydrogenase subunits. Lanes 2 to 5 have HydB C-terminus (608) gene insert, which is 433bp. Ladder is Hyperladder Kb, sizes shown in bp

DNA of the colony on lane 3 was sent off for sequencing. After sequencing confirmation it was used for the Golden Gate cloning and submitted into the iGEM registry.

Restriction site incompatibility, [1000], is with regards to MoClo i.e. Golden Gate cloning. As we used this in downstream cloning experiments the restriction site could not be avoided.

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
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1
Illegal BsaI.rc site found at 369

@@ Line 5: / Line 5: @@
 One of the three subunits that make up FeFe Hydrogenase in Shewanella oneidensis. Specifically this is the HydB subunit which is the central subunit. This basic part has a strep-tag attached to its C-terminus for consequent protein assays. This unit is one out of the three subunits that can be used to make a the entire FeFe hydrogenase construct.
-[[File:T--NRP-UEA-Norwich--Gel_PCR_Colony_FeFe_.jpg|thumb|Figure 1: PCR Colony of FeFe Hydrogenase subunits. Lanes 2 to 5 have HydB C-terminus (608) gene insert, which is 433bp. Ladder is Hyperladder Kb, sizes shown in bp]]
+[[File:T--NRP-UEA-Norwich--Gel_PCR_Colony_FeFe_.jpg|thumb|none|Figure 1: PCR Colony of FeFe Hydrogenase subunits. Lanes 2 to 5 have HydB C-terminus (608) gene insert, which is 433bp. Ladder is Hyperladder Kb, sizes shown in bp]]
 DNA of the colony on lane 3 was sent off for sequencing. After sequencing confirmation it was used for the Golden Gate cloning and submitted into the iGEM registry.
 Restriction site incompatibility, [1000], is with regards to MoClo i.e. Golden Gate cloning. As we used this in downstream cloning experiments the restriction site could not be avoided.