Part:BBa_K3159001
NFX1
1 Usage and Biology
MHC(Major Histocompatibility Complex) Class II cells are crucial in immune cell communication. It informs other cells of the impeding danger so that they can be prepared and build necessary defense against the virus, restraining the virus from spreading. The gene expression of MHC Class II cells are primarily controlled by transcription factors that bind to X and Y boxes. NFX1 is one of the transcriptional factor. This sequence codes for a transcriptional repressor protein, which binds to MHC Class II genes on the highly conserved x-box motif in vitro to regulate gene expression. Malfunctioning of this protein may decrease the sensitivity of MHC Class II cells in their inflammatory immune response induced by Interferon-gamma, shortening the period in which defense can be built against antigens.
2 Plasmid construction
2.1 pLEX-MCS Vector
pLEX is a DNA molecule that is capable of self-replication. This genetic vector is often used in genetic programming to transact the gene of interest into a cell, resulting in an over-expression of the targeted gene. In other words, it’s an indispensable pathway that our gene of interest has to take in order to be expressed in a cell. Thus, constructing plasmids is a necessary step for us to get the protein coded by our inserted gene. In our experiment, the pLEX being used for all targeted gene is the pLEX-MCS Empty Vector. One of the benefits of this artificial vector is that it contains multiple restriction enzyme cutting sites, which is ideal for the design of primers for target gene, transfection, and subsequent transduction in E. coli.
Figure 1: stricture of pLEX-MCS used
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 960
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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