Impact manual intein

 · In comparison, to achieve a similar extent of C-terminal cleavage, it takes ∼16 h at 23°C for the IMPACT system (New England Biolabs, Ipswich, MA, USA) employing SceVma1 and SspDnaB intein (Chong et al., a, b) (IMPACT Manual). IMPACT (Intein Mediated Purification with an Affinity Chitin-binding Tag) is a novel protein purifica-tion system which utilizes the inducible self-cleavage activity of a protein splicing element (termed intein) to separate the target protein from the affinity tag. It distinguishes itself from all other purifi-.  · Thus, the intein-mediated self-cleavage tag was developed and has been commercially available as the IMPACT™ system. However, uncontrolled cleavages .

The IMPACT system, which circumvents the protease problem through DTT-induced intein cleaving, is the third system we have analyzed. Finally, two recently-developed PHB and ELP methods, which allow pH-induced intein cleaving by the ΔI-CM mini intein and virtually eliminate affinity resin costs, are also included for comparison (Figure 1). USB1 US09/, USA USB1 US B1 US B1 US B1 US A US A US A US B1 US B1 US B1 Authority US United States Prior art keywords protein intein target protein precursor target Prior art date Legal status (The legal status is an assumption and is not a legal conclusion. Most inteins (intein endonucleases) possess two conserved LAGLIDADG ( Mutational analysis of active‐site residues in the Mycobacterium leprae RecA intein, a LAGLIDADG homing endonuclease: Asp and Asp are crucial to the double‐stranded DNA cleavage activity whereas Asp is not - Singh - - Protein Science - Wiley Online Library.

IMPACT (Intein Mediated Purification with an Affinity Chitin-binding Tag) is a novel protein purifica-tion system which utilizes the inducible self-cleavage activity of a protein splicing element (termed intein) to separate the target protein from the affinity tag. It distinguishes itself from all other purifi-. The IMPACT (Intein Mediated Purification with an Affinity Chitin-binding Tag) system is a novel protein purification system which utilizes the inducible self-cleavage activity of protein splicing elements (termed inteins) to separate the target protein from the affinity tag (1). It distinguishes itself from all other. The IMPACT (Intein Mediated Purification with an Affinity Chitin-binding Tag) system is a novel protein purification strategy that utilizes the inducible self-cleavage activity of protein splicing elements (termed inteins) to separate the target protein from the affinity tag (1).