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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 INSR ORF mammalian expression plasmid, C-GFPSpark tag||RG80099-ACG|
|Rat INSR ORF mammalian expression plasmid, C-OFPSpark / RFP tag||RG80099-ACR|
|Rat INSR ORF mammalian expression plasmid, C-Flag tag||RG80099-CF|
|Rat INSR ORF mammalian expression plasmid, C-His tag||RG80099-CH|
|Rat INSR ORF mammalian expression plasmid, C-Myc tag||RG80099-CM|
|Rat INSR ORF mammalian expression plasmid, C-HA tag||RG80099-CY|
|Rat INSR ORF mammalian expression plasmid, N-Flag tag||RG80099-NF|
|Rat INSR ORF mammalian expression plasmid, N-His tag||RG80099-NH|
|Rat INSR ORF mammalian expression plasmid, N-Myc tag||RG80099-NM|
|Rat INSR ORF mammalian expression plasmid, N-HA tag||RG80099-NY|
|Rat INSR natural ORF mammalian expression plasmid||RG80099-UT|
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INSR (Insulin receptor), also known as CD220, is a transmembrane receptor that is activated by insulin. INSR belongs to theprotein kinase superfamily, and exists as a tetramer consisting of two alpha subunits and two beta subunits linked by disulfide bonds. The alpha and beta subunits are encoded by a single INSR gene, and the beta subunits pass through the cellular membrane. As the receptor for insulin with tyrosine-protein kinase activity, INSR associates with downstream mediators upon binding to insulin, including IRS1 (insulin receptor substrate 1) and phosphatidylinositol 3'-kinase (PI3K). IRS-1 binding and phosphorylation eventually leads to an increase in the high affinity glucose transporter (Glut4) molecules on the outer membrane of insulin-responsive tissues. INSR isoform long and isoform short are expressed in the peripheral nerve, kidney, liver, striated muscle, fibroblasts and skin, and is found as a hybrid receptor with IGF1R which also binds IGF1 in muscle, heart, kidney, adipose tissue, skeletal muscle, hepatoma, fibrobasts, spleen and placenta. Defects in Insulin Receptor/INSR are the cause of Rabson-Mendenhall syndrome (Mendenhall syndrome), insulin resistance (Ins resistance), leprechaunism (Donohue syndrome), and familial hyperinsulinemic hypoglycemia 5 (HHF5). It may also be associated with noninsulin-dependent diabetes mellitus (NIDDM).