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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 EFNB1 ORF mammalian expression plasmid, C-GFPSpark tag||RG80106-ACG|
|Rat EFNB1 ORF mammalian expression plasmid, C-OFPSpark / RFP tag||RG80106-ACR|
|Rat EFNB1 ORF mammalian expression plasmid, C-Flag tag||RG80106-CF|
|Rat EFNB1 ORF mammalian expression plasmid, C-His tag||RG80106-CH|
|Rat EFNB1 ORF mammalian expression plasmid, C-Myc tag||RG80106-CM|
|Rat EFNB1 ORF mammalian expression plasmid, C-HA tag||RG80106-CY|
|Rat EFNB1 ORF mammalian expression plasmid, N-Flag tag||RG80106-NF|
|Rat EFNB1 ORF mammalian expression plasmid, N-His tag||RG80106-NH|
|Rat EFNB1 ORF mammalian expression plasmid, N-Myc tag||RG80106-NM|
|Rat EFNB1 ORF mammalian expression plasmid, N-HA tag||RG80106-NY|
|Rat EFNB1 natural ORF mammalian expression plasmid||RG80106-UT|
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Ephrin-B1 also known as EFNB1, is a member of the ephrin family. The transmembrane- associated ephrin ligands and their Eph family of receptor tyrosine kinases are expressed by cells of the SVZ. Eph/ephrin interactions are implicated in axon guidance, neural crest cell migration, establishment of segmental boundaries, and formation of angiogenic capillary plexi. Eph receptors and ephrins are divided into two subclasses, A and B, based on binding specificities. Ephrin subclasses are further distinguished by their mode of attachment to the plasma membrane: ephrin-A ligands bind EphA receptors and are anchored to the plasma membrane via a glycosylphosphatidylinositol (GPI) linkage, whereas ephrin-B ligands bind EphB receptors and are anchored via a transmembrane domain. An exception is the EphA4 receptor, which binds both subclasses of ephrins. EphrinB1 and B class Eph receptors provide positional cues required for the normal morphogenesis of skeletal elements. Another malformation, preaxial polydactyly, was exclusively seen in heterozygous females in which expression of the X-linked ephrinB1 gene was mosaic, so that ectopic EphB-ephrinB1 interactions led to restricted cell movements and the bifurcation of digital rays.