A general method of protein purification for recombinant unstructured non-acidic proteins
Por:
Campos, F, Guillen, G, Reyes, JL, Covarrubias, AA
Publicada:
1 nov 2011
Categoría:
Biotechnology
Resumen:
Typical late embryogenesis abundant (LEA) proteins accumulate in response to water deficit imposed by the environment or by plant developmental programs. Because of their physicochemical properties, they can be considered as hydrophilins and as a paradigm of intrinsically unstructured proteins (IUPs) in plants. To study their biophysical and biochemical characteristics large quantities of highly purified protein are required. In this work, we report a fast and simple purification method for non-acidic recombinant LEA proteins that does not need the addition of tags and that preserves their in vitro protective activity. The method is based on the enrichment of the protein of interest by boiling the bacterial protein extract, followed by a differential precipitation with trichloroacetic acid (TCA). Using this procedure we have obtained highly pure recombinant LEA proteins of groups 1, 3, and 4 and one recombinant bacterial hydrophilin. This protocol will facilitate the purification of th
Filiaciones:
Campos, F:
Univ Nacl Autonoma Mexico, Inst Biotecnol, Dept Biol Mol Plantas, Cuernavaca 62210, Morelos, Mexico
Guillen, G:
Univ Nacl Autonoma Mexico, Inst Biotecnol, Dept Biol Mol Plantas, Cuernavaca 62210, Morelos, Mexico
Reyes, JL:
Univ Nacl Autonoma Mexico, Inst Biotecnol, Dept Biol Mol Plantas, Cuernavaca 62210, Morelos, Mexico
Covarrubias, AA:
Univ Nacl Autonoma Mexico, Inst Biotecnol, Dept Biol Mol Plantas, Cuernavaca 62210, Morelos, Mexico
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