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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.
|Mouse MANF ORF mammalian expression plasmid, C-GFPSpark tag||MG50954-ACG|
|Mouse MANF ORF mammalian expression plasmid, C-OFPSpark / RFP tag||MG50954-ACR|
|Mouse MANF ORF mammalian expression plasmid, C-Flag tag||MG50954-CF|
|Mouse MANF ORF mammalian expression plasmid, C-His tag||MG50954-CH|
|Mouse MANF ORF mammalian expression plasmid, C-Myc tag||MG50954-CM|
|Mouse MANF ORF mammalian expression plasmid, C-HA tag||MG50954-CY|
|Mouse MANF ORF mammalian expression plasmid, N-Flag tag||MG50954-NF|
|Mouse MANF ORF mammalian expression plasmid, N-His tag||MG50954-NH|
|Mouse MANF ORF mammalian expression plasmid, N-Myc tag||MG50954-NM|
|Mouse MANF ORF mammalian expression plasmid, N-HA tag||MG50954-NY|
|Mouse MANF natural ORF mammalian expression plasmid||MG50954-UT|
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Mesencephalic astrocyte-derived neurotrophic factor, also known as Protein ARMET, Arginine-rich protein, MANF and ARMET, is a secreted protein which belongs to the ARMET family. ARMET selectively promotes the survival of dopaminergic neurons of the ventral mid-brain. It modulates GABAergic transmission to the dopaminergic neurons of the substantia nigra. ARMET enhances spontaneous, as well as evoked, GABAergic inhibitory postsynaptic currents in dopaminergic neurons. ARMET inhibits cell proliferation and endoplasmic reticulum (ER) stress-induced cell death. The N-terminal region of ARMET may be responsible for neurotrophic activity while the C-terminal region may play a role in the ER stress response. MANF reduces endoplasmic reticulum (ER) stress and has neurotrophic effects on dopaminergic neurons. Intracortical delivery of recombinant MANF protein protects tissue from ischemic brain injury. MANF has been described as a survival factor for dopaminergic neurons. MANF expression was widespread in the nervous system and non-neuronal tissues. In the brain, relatively high MANF levels were detected in the cerebral cortex, hippocampus and cerebellar Purkinje cells. The widespread expression of MANF together with its evolutionary conserved nature and regulation by brain insults suggest that it has important functions both under normal and pathological conditions in many tissue types.