Difference between revisions of "Part:BBa K1714003"

Revision as of 23:26, 18 September 2015

pLac-RBS B0034-His10-TEV-thanatin-double terminator

This part is designed for thanatin expression system regulated by pLac and decreased the toxicity by adding His10.

By adding IPTG, this part express His10 - TE - thanatin.
The His10 is also used for purification. After purification, they can be cleaved His10 and reactivated by TEV protease.

Even we subcultured many colonies from LB plate to liquid culture, the number of correctly subcultured products were few. We expected this result could be caused by thanatin's strong toxicity. Expect naturally modified cells, losing thanatin expression systems were not successed. It's very hard to survive with thanatin expression system even their toxicity was decreased by adding His10 and to get ideal sequences. We recommend you check whether this part's sequence is ideal sequence or not.

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
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 287
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1714003 short</partinfo>
-This part is designed for thanatin expression system regulated by pLac and decreased the toxicity by adding His10. The His10 is also used for purification. After purification, they can be cleaved his10 and reactivated by TEV protease. By adding IPTG, this part express His10-TE-thanatin. However, even we sequenced this part, the sequence was awfully different from that as we expected. In addition, even we subcultured many colonies from LB plate to liquid culture, the number of correctly subcultured products were so few. We thought because of thanatin's strong toxicity, it's very hard to survive with thanatin expression system even their toxicity was decreased by adding His10. So only host cells that lost thanatin expression systems were successfully cultured. So we recommend you not to use this part because this part's sequence may be different from the sequence on this page.
+This part is designed for thanatin expression system regulated by pLac and decreased the toxicity by adding His10.
+ By adding IPTG, this part express His10 - TE - thanatin.
+ The His10 is also used for purification. After purification, they can be cleaved His10 and reactivated by TEV protease.
+Even we subcultured many colonies from LB plate to liquid culture, the number of correctly subcultured products were few.
+We expected this result could be caused by thanatin's strong toxicity. Expect naturally modified cells, losing thanatin expression systems were not successed.
+It's very hard to survive with thanatin expression system even their toxicity was decreased by adding His10 and to get ideal sequences.
+We recommend you check whether this part's sequence is ideal sequence or not.
 <!-- Add more about the biology of this part here