Difference between revisions of "Part:BBa K2403003"
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===Usage and Biology=== | ===Usage and Biology=== | ||
+ | In eukaryotic cells, it is known that RNA is transcribed in the nucleus and then exported to the cytoplasm, a process mediated by nuclear export factors specific to each type of RNA. In the case of single stranded mRNA, the TAP / p15 complex binds and exports it to the cytoplasm. However, artificial RNAs designed for synthetic biology applications are often highly structured RNA which are not recognized by TAP / p15, and therefore a different pathway is required to export such target RNAs to the cytoplasm. | ||
+ | |||
+ | In order to solve this problem, we used the Rev protein derived from HIV-1 which binds to and exports RNA containing the cis-acting RRE sequence (Rev Response Element). By fusing the RRE sequence to an RNA of interest and co-expressing Rev protein, complex RNA is exported. | ||
+ | |||
+ | ===Characterization=== | ||
+ | In order to demonstrate the function of the Rev protein and RRE, we fused an RRE [1] to U6 snRNA, which has a complex structure and is not transported by the TAP / p15 pathway. In Xenopus oocytes, we ascertained whether this RNA was exported from the nucleus in a Rev-dependent manner. | ||
+ | As shown in the figure, the U6-RRE RNA itself remains in the nucleus even 60 minutes after injection. On the other hand, the same RNA was exported to the cytoplasm when Rev was co-injected. From this data, we showed that a combination of RRE and Rev allowed target RNAs to be exported to the cytoplasm. | ||
+ | |||
+ | <!-- (以下Rev アフリカツメガエルinjection result、and analysis fig) --> | ||
+ | |||
+ | ===Reference=== | ||
+ | [1]https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4041468/pdf/gku304.pdf | ||
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Revision as of 17:49, 31 October 2017
TDH3 promoter>EGFP protein
We expressed this gene in Saccharomyces Cerevisiae and feed it to B.xylophilus.
Reference
[1]https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4041468/pdf/gku304.pdf
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]