Part:BBa_K929001
AID without NES, with NLS and Kozak sequence
| AID without NES, with NLS and Kozak sequence | |
|---|---|
| |
| BioBrick Nr. | BBa_929001 |
| RFC standard | RFC 10 |
| Requirement | pSB1C3 |
| Source | existing parts: |
| Submitted by | [http://2012.igem.org/Team:Potsdam_Bioware Potsdam_Bioware2012] |
The BioBrick wtAID is an improved version of the existing AID BioBrick (BBa_K103001). It has no NES(Nuclear Export Sequence) sequence and has NLS (Nuclear Localization Sequence) sequence and Kozak sequence added
The AID motif is naturally terminated with the Nuclear Export Sequence (NES) that causes the protein to translocate from the nucleus to the cytoplasm. Additionally, upstream, dysfunctional Nuclear Localization Sequence (NLS) is located. Due to the fact that AID mutates the actively transcribed single stranded DNA, it is supposed that the direction of the enzyme to the inside of the nucleus would improve the mutation rate.
AID:
AID is known to be responsible for somatic hypermutation and the class-switch recombination of immunoglobulin in B cells. This enzyme of 28 kDa originally occurs in B cells but does also show activity after transfection into CHO cells. AID induces the deamination of cytidine to uridine at actively transcribed single strand DNA. The replacement of cytidine by uridine leads to a mismatch during DNA replication and integrates a single base substitution predominantly in the immunoglobulin genes.
Functional NLS sequence:
This part of the BioBrick directs the expressed protein into the nucleus, where it can mutate stronger.
Kozak sequence
Kozak consensus sequence is added upstream of the AID mutant to express the protein stronger.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 579
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 108
Illegal SapI site found at 209
| None |