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.
|Human PI3 ORF mammalian expression plasmid, C-GFPSpark tag||HG12187-ACG|
|Human PI3 ORF mammalian expression plasmid, C-OFPSpark / RFP tag||HG12187-ACR|
|Human PI3 ORF mammalian expression plasmid, C-Flag tag||HG12187-CF|
|Human PI3 ORF mammalian expression plasmid, C-His tag||HG12187-CH|
|Human PI3 ORF mammalian expression plasmid, C-Myc tag||HG12187-CM|
|Human PI3 ORF mammalian expression plasmid, C-HA tag||HG12187-CY|
|Human PI3 ORF mammalian expression plasmid, N-Flag tag||HG12187-NF|
|Human PI3 ORF mammalian expression plasmid, N-His tag||HG12187-NH|
|Human PI3 ORF mammalian expression plasmid, N-Myc tag||HG12187-NM|
|Human PI3 ORF mammalian expression plasmid, N-HA tag||HG12187-NY|
|Human PI3 natural ORF mammalian expression plasmid||HG12187-UT|
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Elafin, also known as Elastase-specific inhibitor, Peptidase inhibitor 3, Protease inhibitor WAP3, Skin-derived antileukoproteinase, WAP four-disulfide core domain protein 14, PI3, WAP3 and WFDC14, is a secreted protein which contains one WAP domain. Elafin / PI3 consists of two domains: the transglutaminase substrate domain (cementoin moiety) and the elastase inhibitor domain. The transglutaminase substrate domain at the N-terminus serves as an anchor to localize elafin covalently to specific sites on extracellular matrix proteins. The serine anti-protease Elafin / PI3 is expressed by monocytes, alveolar macrophages, neutrophils, and at mucosal surfaces and possesses antimicrobial activity. It is also known to reduce lipopolysaccharide-induced neutrophil influx into murine alveoli as well as to abrogate lipopolysaccharide-induced production of matrix metalloprotease 9, macrophage inhibitory protein 2, and tumor necrosis factor-alpha by as-yet unidentified mechanisms. Elafin / PI3 is a neutrophil serine protease inhibitor expressed in lung and displaying anti-inflammatory and anti-bacterial properties. Elafin / PI3 is a neutrophil and pancreatic elastase-specific inhibitor of skin. It may prevent elastase-mediated tissue proteolysis. Elafin / PI3 will regulate proteolytic enzymes during menstruation and will contribute to the innate defense against uterine infection.