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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 GDF10 ORF mammalian expression plasmid, C-GFPSpark tag||RG80130-ACG|
|Rat GDF10 ORF mammalian expression plasmid, C-OFPSpark / RFP tag||RG80130-ACR|
|Rat GDF10 ORF mammalian expression plasmid, C-Flag tag||RG80130-CF|
|Rat GDF10 ORF mammalian expression plasmid, C-His tag||RG80130-CH|
|Rat GDF10 ORF mammalian expression plasmid, C-Myc tag||RG80130-CM|
|Rat GDF10 ORF mammalian expression plasmid, C-HA tag||RG80130-CY|
|Rat GDF10 ORF mammalian expression plasmid, N-Flag tag||RG80130-NF|
|Rat GDF10 ORF mammalian expression plasmid, N-His tag||RG80130-NH|
|Rat GDF10 ORF mammalian expression plasmid, N-Myc tag||RG80130-NM|
|Rat GDF10 ORF mammalian expression plasmid, N-HA tag||RG80130-NY|
|Rat GDF10 natural ORF mammalian expression plasmid||RG80130-UT|
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BMP-3b / GDF-10 is a member of the bone morphogenetic protein (BMP) family and the TGF-beta superfamily. This group of proteins is characterized by a polybasic proteolytic processing site which is cleaved to produce a mature protein containing seven conserved cysteine residues. The members of this family are regulators of cell growth and differentiation in both embryonic and adult tissues. Studies in mice suggest that the protein encoded by this gene plays a role in skeletal morphogenesis. In the bone morphogenetic cascade, cartilage differentiation, hypertrophy, and cell death are followed by bone formation. In this regard, all BMPs are cartilage morphogenetic proteins since cartilage is formed first. An overexpression or dysregulation of BMP pathways may lead to heterotopic bone formation or fibrodysplasia ossificans progressiva (FOP). BMPs have been implicated in FOP. The pioneering work of Sakou has implicated BMP-3b / GDF-10 in ossification of the posterior longitudinal ligament of the spine in Japanese patients. The BMP-specific antagonists such as noggin or chordin might be used therapeutically in clinical conditions with pathologically excessive bone formation. The osteoinductive capacity of BMPs has been demonstrated in preclinical models, and the efficacy of BMPs for the treatment of orthopaedic patients is now being evaluated in clinical trials. It was suggested that further progress in the clinical application of the BMP-3b / GDF-10 will depend upon the development of carriers with ideal release kinetics for the delivery of the BMPs.