Coding

Part:BBa_K4414003

Designed by: Yue Yin   Group: iGEM22_NUDT_CHINA   (2022-09-22)
Revision as of 13:47, 9 October 2022 by Bingshun (Talk | contribs)


NES

This nuclear export signal(NES)directs the protein out of the nucleus.

Usage and Biology

Regulating the output of many proteins from the nucleus depends on the presence of a nuclear export signal(NES) consisting of leucine-rich amino acids.[1][2]. The shuttle receptor that appears to bind to the NES sequence and function in the protein output from the nucleus to the cytoplasm is CRM1 (chromosomal region maintenance)/export protein 1.[3] CRM1 binds to Ran GTPase and interacts with the nucleoporous components to translocate NES-containing proteins to the cytoplasm.


Sequecing

The plasmid was sequenced correct.


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]



Functional characterization

We can connect NES to EGFP and observe the location of fluorescent proteins to verify the function of NES.In order to test the effect of NES on background value reduction, we compared the SEAP characterization effect of components without NES with those adding NES.


Reference

[1].Fischer, U., Huber, J., Boelens, W. C., Mattaj, I. W., & Lührmann, R. (1995). The HIV-1 Rev activation domain is a nuclear export signal that accesses an export pathway used by specific cellular RNAs. Cell, 82(3), 475–483. https://doi.org/10.1016/0092-8674(95)90436-0 [2].Fornerod, M., Ohno, M., Yoshida, M., & Mattaj, I. W. (1997). CRM1 is an export receptor for leucine-rich nuclear export signals. Cell, 90(6), 1051–1060. https://doi.org/10.1016/s0092-8674(00)80371-2 [3].Richards, S. A., Carey, K. L., & Macara, I. G. (1997). Requirement of guanosine triphosphate-bound ran for signal-mediated nuclear protein export. Science (New York, N.Y.), 276(5320), 1842–1844. https://doi.org/10.1126/science.276.5320.1842

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