BIOL 4087 Chapter : BioChem Questions
Document Summary
From moodle: what characteristic of the polyunsaturated fatty acids that allows peroxidation by. Please describe in detail the procedure for expression and purification of 15-lipoxygenase-2. In order amplify 15-lox-2 expression, the gene was coexpressed in rossetta 2 cells in a pet duet-1 with. The genes were then amplified from e. coli dna and cloned behind promoter. 2 in pet duet-1 by digesting both vectors and inserting them. The human 15-lox-2 cdna obtained from the atcc was amplified, by digesting the vector with xbai and hindiii, transforming primers containing spei and hindiii sites. The rossetta 2 cells were then allowed to grow, expressing the gene and forming 15-lox-2. For purification, cells are lysed using both invitrogen and problock 2d, a protease inhibitor. Dnase i, insoluble cell material was centrifuged and removed while the supernate was mixed with imidazole buffer and filtered.
