Difference between revisions of "Part:BBa K929000"
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Revision as of 23:50, 25 September 2012
AID with CMV promoter and hGH-polyadenylation signal sequence
General information
CMV_wtAID_pA | |
---|---|
BioBrick Nr. | BBa_K929000 |
RFC standard | RFC 10 |
Requirement | pSB1C3 |
Source | existing parts:(BBa_K103001;BBa_I712004; BBa_K404108) |
Submitted by | [http://2012.igem.org/Team:Potsdam_Bioware Potsdam_Bioware2012] |
The BioBrick "AID with CMV promoter and hGH-polyadenylation signal sequence" (BBa_K929000) is an extended version of the existing AID BioBrick (BBa_K103001). It is built of 3 parts: CMV promoter (BBa_I712004), AID BBa_K103001)and hGH polyadenylation signal sequence (BBa_K404108).
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.
CMV promoter:
CMV is an immediate-early Cytomegalovirus promoter for high-level expression. The CMV promoter is commonly used due to its very strong activity and effectivity in a broad range of cell types. The BioBrick is therefore improved via addition of the strong promoter.
hGH polyadenylation signal sequence:
Polyadenylation is a significant part for the translation and stability of mRNA. In eukaryotes, it is part of the process that produces mature messenger RNA (mRNA) for translation. It, therefore, forms part of the larger process of gene expression. hGH terminator gives a signal to start polyadenylation in the translation process.
Characterization
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1661
Illegal BsaI.rc site found at 740
Illegal SapI site found at 841