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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 HRG ORF mammalian expression plasmid, C-GFPSpark tag||RG81643-ACG|
|Rat HRG ORF mammalian expression plasmid, C-OFPSpark / RFP tag||RG81643-ACR|
|Rat HRG ORF mammalian expression plasmid, N-GFPSpark tag||RG81643-ANG|
|Rat HRG ORF mammalian expression plasmid, N-OFPSpark / RFP tag||RG81643-ANR|
|Rat HRG ORF mammalian expression plasmid, C-Flag tag||RG81643-CF|
|Rat HRG ORF mammalian expression plasmid, C-His tag||RG81643-CH|
|Rat HRG ORF mammalian expression plasmid, C-Myc tag||RG81643-CM|
|Rat HRG ORF mammalian expression plasmid, C-HA tag||RG81643-CY|
|Rat HRG Gene cDNA clone plasmid||RG81643-G|
|Rat HRG ORF mammalian expression plasmid, N-Flag tag||RG81643-NF|
|Rat HRG ORF mammalian expression plasmid, N-His tag||RG81643-NH|
|Rat HRG ORF mammalian expression plasmid, N-Myc tag||RG81643-NM|
|Rat HRG ORF mammalian expression plasmid, N-HA tag||RG81643-NY|
|Rat HRG natural ORF mammalian expression plasmid||RG81643-UT|
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Histidine-rich glycoprotein, also known as HRG and HPRG, is a glycoprotein located in plasma and platelets, and contains an unusually large amount of histidine and proline. In human, five distinct domains are recognized in the mature HPRG molecule. There are two N-terminal cystatin-like modules (aa 19 - 254) and one His-Pro-rich region (aa 350 - 497) that is flanked by two Pro-rich segments (aa 276 - 321 and 498 - 525). The His-Pro-rich region contains 10 tandem repeats with an HHPHG motif, and the N- and C-termini are linked by a disulfide bond. The specific functions of HRG remain unclear, but it is known that the protein binds heme, dyes and divalent metal ions. It inhibits rosette formation and interacts with heparin, thrombospondin and plasminogen. Two of the protein's effects, the inhibition of fibrinolysis and the reduction of inhibition of coagulation, indicate a potential prothrombotic effect. HPRG is evolutionarily, functionally and structurally related to cleaved high molecular weight kininogen (HKa), an anti-angiogenic polypeptide that stimulates apoptosis of proliferating endothelial cells through binding to cell-surface tropomyosin. The antiangiogenic activity of the multidomain plasma protein HPRG is localized to its histidine-proline-rich (H/P) domain and has recently been shown to be mediated, at least partially, through binding to cell-surface tropomyosin in fibroblast growth factor-2-activated endothelial cells.