Difference between revisions of "Part:BBa K3402008"

Latest revision as of 23:42, 19 October 2020

NLS

Because of the high molecular weight of Cas9 nuclease which cannot freely shuttle through the nuclear pores, we have to add a nuclear localization sequence(NLS) to guide it through the nucleus.

Usage and Biology

NLS was connected to the 3' end of the yeGFP(BBa_K3402000) gene to construct the plasmid, then we transferred the plasmid to observe the distribution of green fluorescence. If the fluorescent region was concentrated in the nucleus, this sequence could be used for the subsequent establishment of the CRISPR/Cas9 system.

Fig.1 Without the NLS (nuclear localization signal peptide)

Fig.2 With the NLS (nuclear localization signal peptide)

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
COMPATIBLE WITH RFC[1000]

@@ Line 9: / Line 9: @@
 <i>NLS</i> was connected to the 3' end of the <i>yeGFP</i>([https://parts.igem.org/Part:BBa_K3402000# BBa_K3402000]) gene to construct the plasmid, then we transferred the plasmid to observe the distribution of green fluorescence. If the fluorescent region was concentrated in the nucleus, this sequence could be used for the subsequent establishment of the CRISPR/Cas9 system.
-[[Image:Without NLS.png|500px|thumb|center|Without the <i>NLS</i> (nuclear localization signal peptide)]]
+[[Image:Without NLS.png|500px|thumb|center|Fig.1 Without the <i>NLS</i> (nuclear localization signal peptide)]]
-[[Image:With NLS.png|500px|thumb|center|With the <i>NLS</i> (nuclear localization signal peptide)]]
+[[Image:With NLS.png|500px|thumb|center|Fig.2 With the <i>NLS</i> (nuclear localization signal peptide)]]
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