Difference between revisions of "Part:BBa K2381012"

Revision as of 00:06, 2 November 2017

3'-OriC gRNA

It’s a gRNA sequence containing 20bp targeting at oriC and a full-length gRNA scaffold. The sequence is central to our project and is related with all the system built this year.
The targeting sequence of this part is acquired from reference paper.

Its function after J23119 promoter, BBa_K2381024, is proved by OD measurement after cotransformed with plasmid containing dCas9. Its effect is even better than rapamycin, a well-used chemical molecule for replication inhibition.

Further investigation is conducted by qPCR and flow cytometric.

Its function after light-induced promoter, BBa_K2381023, is proved by OD measurement.

References

Wiktor, J., Lesterlin, C., Sherratt, D. J., & Dekker, C. (2016). CRISPR-mediated control of the bacterial initiation of replication. Nucleic Acids Res, 44(8), 3801-3810.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 4
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 6: / Line 6: @@
 It’s a gRNA sequence containing 20bp targeting at <i>oriC</i> and a full-length gRNA scaffold. The sequence is central to our project and is related with all the system built this year.<br/>
 The targeting sequence of this part is acquired from reference paper. </p>
-[[File:T--HZAU-China--gRNA_function.png|200px|thumb|left|Its function after J23119 promoter, <partinfo>BBa_K2381024</partinfo>, is proved by OD measurement after cotransformed with plasmid containing dCas9.We can observe tight repression on cell growth. ]]
+[[File:T--HZAU-China--gRNA_function.png|200px|thumb|left|Its function after J23119 promoter, <partinfo>BBa_K2381024</partinfo>, is proved by OD measurement after cotransformed with plasmid containing dCas9. Its effect is even better than rapamycin, a well-used chemical molecule for replication inhibition. ]]
 [[File:T--HZAU-China--gRNA_qPCR.png|200px|thumb|left|Further investigation is conducted by qPCR and flow cytometric.]]
 [[File:T--HZAU-China--light_gRNA.png|200px|thumb|left|Its function after light-induced promoter, <partinfo>BBa_K2381023</partinfo>, is proved by OD measurement.]]