pEcoli-6xHN-GFPuv is a control vector that encodes a GFPuv fusion protein containing an N-terminal 6xHN tag (composed of 6 repeating His-Asn subunits) and an enterokinase cleavage (EK) site. High-level, IPTG-inducible expression of the fusion is driven by the T7 lac hybrid promoter (PT7 lac), which consists of the strong T7 promoter (1–3) and the lac operator (4). Basal expression of the fusion is repressed by the Lac repressor (lacI), which binds to the lac operator, preventing expression from the promoter in the absence of IPTG. High-level, IPTG-inducible expression of 6xHN-tagged GFPuv is driven by the T7 promoter in the presence of T7 RNA polymerase.

The vector also contains an ampicillin resistance gene (Ampr) and a pBR322 origin of replication, which maintains the vector at a low-copy-number to further reduce basal levels of the fusion.

載體應(yīng)用

pEcoli-6xHN-GFPuv is included with Clontech’s pEcoli Expression System and pEcoli Linear Expression System (Cat. Nos. 631417 & 631418, respectively). GFPuv is a green fluorescent protein variant optimized for maximal fluorescence when excited by UV light. The vector allows the use of GFPuv fluorescence (excitation and emission maxima at 395 nm and 509 nm, respectively) to monitor protein expression and purification.

His-tagged proteins, including 6xHN-GFPuv, can be readily detected with either our Universal His Western Blot Kit (Cat No. 635633) or our 6xHN Polyclonal Antibody (631207). Exceptionally pure his-tagged proteins can be obtained with our TALON Co resins (Cat. Nos. 635501–635507, 635509 & 635510) and columns (Cat. Nos. 635601–635603 and 635606). For routine use, we have a variety of high capacity His60 Ni resins available (Cat. Nos. 635659–635664). In addition, an assortment of starter kits containing your choice of TALON Co or His60 Ni resin, as well as pEcoli-Nterm/-Cterm 6xHN circular or linear vectors (Cat Nos. 631416, and 631419–631427) are also available.