There is disclosed a method for providing for the coexpression of N-myristoyltransferase and a protein substrate for said N-myristoyltransferase in E. coli comprising introducing into E. coli a dual plasmid system comprising (A) the isopropyl-β-D-thiogalactopyranoside-inducible tac promoter, the g10-L ribosome binding site, a NMT gene, the kanamycin resistance gene and the p15A origin of replication in operable sequence and (B) the recA promoter, the g10-L ribosome binding site, a mammalian gene, the ampicillin resistance gene and the Col E1 origin of replication in operable sequence. This allows production of mammalian N-myristoylproteins or proteins containing covalently linked analogs of myristate with altered physical-chemical properties.
大肠杆菌的双质粒系统包括:(A) 异丙基-β-D-
硫代
吡喃半
乳糖苷诱导的 tac 启动子、g10-L
核糖体结合位点、N
MT 基因、
卡那霉素抗性
基因和 p15A 复制源的可操作序列;(B) recA 启动子、g10-L
核糖体结合位点、哺乳动物
基因、
氨苄西林抗性
基因和 Col E1 复制源的可操作序列。这样就能生产出哺乳动物的 N-肉豆蔻酰蛋白质或含有共价连接的
肉豆蔻酸类似物的蛋白质,其物理
化学性质也会发生改变。