Part:BBa_K2624002:Design
Nuclear localized NS5B
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 492
Illegal BglII site found at 1340 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 537
Design Notes
The 20 amino acid residues in the C termial have been removed in the part sequence, ensuring cytoplasmic RdRp activity and no anchoring to the endoplasmic reticulum in human cells. Then it is fused with a nuclear localization signal (NLS) to direct the polymerase into the nucleus.
Source
We have one in stock at our lab, and it's consistent with the ducumented BBa_K1442100 of iGEM14_Warwick. It is supposed to have been derived from the Hepatitis C virus con1 strain, but we didn't refer to any public database.
References
[1]Moradpour D, Brass V, Gosert R, et al. Hepatitis C: molecular virology and antiviral targets[J]. Trends in molecular medicine, 2002, 8(10): 476-482.
[2]Brass V, Gouttenoire J, Wahl A, et al. Hepatitis C virus RNA replication requires a conserved structural motif within the transmembrane domain of the NS5B RNA-dependent RNA polymerase[J]. Journal of virology, 2010, 84(21): 11580-11584.
[3]Vo N V, Tuler J R, Lai M M C. Enzymatic characterization of the full-length and C-terminally truncated hepatitis C virus RNA polymerases: function of the last 21 amino acids of the C terminus in template binding and RNA synthesis[J]. Biochemistry, 2004, 43(32): 10579-10591.
[4]Lee K J, Choi J, Ou J, et al. The C-terminal transmembrane domain of hepatitis C virus (HCV) RNA polymerase is essential for HCV replication in vivo[J]. Journal of virology, 2004, 78(7): 3797-3802.
[5]Kao C C, Yang X, Kline A, et al. Template requirements for RNA synthesis by a recombinant hepatitis C virus RNA-dependent RNA polymerase[J]. Journal of virology, 2000, 74(23): 11121-11128.