title: Purification of green fluorescent protein overexpressed by a mutant recombinant Escherichia coli.
abstract: Green fluorescent protein was purified from sonicated recombinant Escherichia coli and its mutant obtained after exposure to UV light. The latter overexpresses green fluorescent protein. The two-step procedure consisted of a two-phase aqueous extraction with PEG/salt and precipitation of the proteins from PEG phase by free Zn2+. The recoveries of green fluorescent protein were 73 and 83% in the cases of recombinant E. coli and its mutant, respectively. The corresponding fold purifications were 24 and 9, respectively. In both cases, the purified protein showed a single band on SDS-PAGE corresponding to 28 kDa.
authors.affiliation: Chemistry Department, Indian Institute of Technology, Delhi Hauz Khas, New Delhi 110016, India.
authors.forename: Sulakshana
authors.initials: S
authors.lastname: Jain
authors.forename: Sunita
authors.initials: S
authors.lastname: Teotia
authors.forename: Munishwar N
authors.initials: MN
authors.lastname: Gupta
mesh_terms: D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification*; D009154:Mutation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins