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.
|人 KIRREL1/NEPH1 基因ORF全长cDNA克隆(表达载体), C-GFPSpark 标签||HG15752-ACG|
|人 KIRREL1/NEPH1 基因ORF全长cDNA克隆(表达载体), C-OFPSpark 标签||HG15752-ACR|
|人 KIRREL1/NEPH1 基因ORF全长cDNA克隆(表达载体), C-Flag 标签||HG15752-CF|
|人 KIRREL1/NEPH1 基因ORF全长cDNA克隆(表达载体), C-His 标签||HG15752-CH|
|人 KIRREL1/NEPH1 基因ORF全长cDNA克隆(表达载体), C-Myc 标签||HG15752-CM|
|人 KIRREL1/NEPH1 基因ORF全长cDNA克隆(表达载体), C-HA 标签||HG15752-CY|
|人 KIRREL1/NEPH1 基因ORF全长cDNA克隆(表达载体), N-Flag 标签||HG15752-NF|
|人 KIRREL1/NEPH1 基因ORF全长cDNA克隆(表达载体), N-His 标签||HG15752-NH|
|人 KIRREL1/NEPH1 基因ORF全长cDNA克隆(表达载体), N-Myc 标签||HG15752-NM|
|人 KIRREL1/NEPH1 基因ORF全长cDNA克隆(表达载体), N-HA 标签||HG15752-NY|
|人 KIRREL1/NEPH1 基因ORF全长cDNA克隆(表达载体)||HG15752-UT|
NEPH1 (KIRREL1) belongs to a family of three closely related transmembrane proteins of the Ig superfamily with a structure similar to that of nephrin. All three Neph proteins share a conserved podocin-binding motif; mutation of a centrally located tyrosine residue dramatically lowers the affinity of Neph1 for podocin. Neph1 triggers AP-1 activation similarly to nephrin but requires the presence of Tec family kinases for efficient transactivation. Neph1 consists of a signal peptide, five Ig-like C2-type domains with the middle domain overlapping with a PKD-like domain, an RGD sequence, a transmembrane domain and a cytoplasmic tail, which is expressed in slit diaphragm domains of podocytes and in vertebrate and invertebrate nervous systems. Neph1 is abundantly expressed in the kidney, specifically expressed in podocytes of kidney glomeruli, and plays a significant role in the normal development and function of the glomerular permeability. Neph1 interacts with nephrin in vitro and in vivo, and able to stimulate transcriptional activation in a model system, such as the activation the transcription factor AP-1 via the stimulation of a MAPK module. Neph1 is crucial for the integrity of the slit diaphragm, as Neph1 gene knockout mice results in effacement of glomerular podocytes, heavy proteinuria, and early postnatal death.