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.
|Canine IL18 ORF mammalian expression plasmid, C-GFPSpark tag||DG70020-ACG|
|Canine IL18 ORF mammalian expression plasmid, C-OFPSpark / RFP tag||DG70020-ACR|
|Canine IL18 ORF mammalian expression plasmid, C-Flag tag||DG70020-CF|
|Canine IL18 ORF mammalian expression plasmid, C-His tag||DG70020-CH|
|Canine IL18 ORF mammalian expression plasmid, C-Myc tag||DG70020-CM|
|Canine IL18 ORF mammalian expression plasmid, C-HA tag||DG70020-CY|
|Canine IL18 ORF mammalian expression plasmid, N-Flag tag||DG70020-NF|
|Canine IL18 ORF mammalian expression plasmid, N-His tag||DG70020-NH|
|Canine IL18 ORF mammalian expression plasmid, N-Myc tag||DG70020-NM|
|Canine IL18 ORF mammalian expression plasmid, N-HA tag||DG70020-NY|
|Canine IL18 natural ORF mammalian expression plasmid||DG70020-UT|
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Interleukin-18 (IL-18, also known as interferon-gamma inducing factor) is a proinflammatory cytokine that belongs to the IL-1 superfamily and is produced by macrophages and other cells. This cytokine can induce the IFN-gamma production of T cells. The combination of IL-18 and IL12 has been shown to inhibit IL4 dependent IgE and IgG1 production, and enhance IgG2a production of B cells. IL-18 binding protein (IL18BP) can specifically interact with this cytokine, and thus negatively regulate its biological activity. IL-18 is an IL-1−like cytokine that requires cleavage with caspase-1 to become active, was found to increase IgE production in a CD4+ T cells-, IL-4− and STAT6−dependent fashion. IL-18 and T cell receptor−mediated stimulation could induce naïve CD4+ T cells to develop into IL-4−producing cells in vitro. Thus, caspase-1 and IL-18 may be critical in regulation of IgE production in vivo, providing a potential therapeutic target for allergic disorders. IL-18 production in primary synovial cultures and purified synovial fibroblasts was, in turn, upregulated by TNF-α and IL-1β, suggesting that monokine expression can feed back to promote Th1 cell development in synovial membrane. Besides, synergistic combinations of IL-18, IL-12, and IL-15 may be of importance in sustaining both Th1 responses and monokine production in RA.