Difference between revisions of "Part:BBa K1714003"
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<partinfo>BBa_K1714003 short</partinfo> | <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 | + | This part is designed for thanatin expression system regulated by pLac and decreased the toxicity by adding His10. |
+ | By adding IPTG, this part expresses 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 | <!-- Add more about the biology of this part here |
Latest revision as of 23:45, 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 expresses 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 287
- 1000COMPATIBLE WITH RFC[1000]