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The pGEM-T is 3kb in length, and contains the amplicin resistance gene, conferring selection of the plasmid in E. coli, and the ori site which is the bacterial origin of replication. The plasmid has multiple cloning sites as shown below. The coding sequence was inserted by TA cloning. Many E. coli strains are suitable for the propagation of this vector including JM109, DH5α and TOP10.
The coding sequence can be easily obtained by digesting the vector with proper restriction enzyme(s). The coding sequence can also be amplified by PCR with M13 primers, or primer pair SP6 and T7.
|Rat KIT ORF mammalian expression plasmid, C-GFPSpark tag||RG80321-ACG|
|Rat KIT ORF mammalian expression plasmid, C-OFPSpark / RFP tag||RG80321-ACR|
|Rat KIT ORF mammalian expression plasmid, C-Flag tag||RG80321-CF|
|Rat KIT ORF mammalian expression plasmid, C-His tag||RG80321-CH|
|Rat KIT ORF mammalian expression plasmid, C-Myc tag||RG80321-CM|
|Rat KIT ORF mammalian expression plasmid, C-HA tag||RG80321-CY|
|Rat KIT ORF mammalian expression plasmid, N-Flag tag||RG80321-NF|
|Rat KIT ORF mammalian expression plasmid, N-His tag||RG80321-NH|
|Rat KIT ORF mammalian expression plasmid, N-Myc tag||RG80321-NM|
|Rat KIT ORF mammalian expression plasmid, N-HA tag||RG80321-NY|
|Rat KIT natural ORF mammalian expression plasmid||RG80321-UT|
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C-Kit is a type 3 transmembrane receptor for MGF (mast cell growth factor, also known as stem cell factor). c-Kit contains 5 Ig-like C2-type (immunoglobulin-like) domains.and 1 protein kinase domain. It belongs to the protein kinase superfamily, tyr protein kinase family and CSF-1/PDGF receptor subfamily. C-Kit contains 5 Ig-like C2-type (immunoglobulin-like) domains and 1 protein kinase domain. C-Kit has a tyrosine-protein kinase activity. Binding of the ligands leads to the autophosphorylation of KIT and its association with substrates such as phosphatidylinositol 3-kinase. Antibodies to c-Kit are widely used in immunohistochemistry to help distinguish particular types of tumour in histological tissue sections. It is used primarily in the diagnosis of GISTs. In GISTs, c-Kit staining is typically cytoplasmic, with stronger accentuation along the cell membranes. C-Kit antibodies can also be used in the diagnosis of mast cell tumours and in distinguishing seminomas from embryonal carcinomas. Mutations in c-Kit gene are associated with gastrointestinal stromal tumors, mast cell disease, acute myelogenous lukemia, and piebaldism. Defects in KIT are a cause of acute myelogenous leukemia (AML). AML is a malignant disease in which hematopoietic precursors are arrested in an early stage of development. Note=Somatic mutations that lead to constitutive activation of KIT are detected in AML patients.