2011
DOI: 10.1371/journal.pone.0020176
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Expression and Purification of Recombinant Hemoglobin in Escherichia coli

Abstract: BackgroundRecombinant DNA technologies have played a pivotal role in the elucidation of structure-function relationships in hemoglobin (Hb) and other globin proteins. Here we describe the development of a plasmid expression system to synthesize recombinant Hbs in Escherichia coli, and we describe a protocol for expressing Hbs with low intrinsic solubilities. Since the α- and β-chain Hbs of different species span a broad range of solubilities, experimental protocols that have been optimized for expressing recom… Show more

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Cited by 52 publications
(64 citation statements)
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“…The twolocus HBB haplotype that increases in frequency as a function of altitude (d 1 d 1 ) is associated with increased hemoglobinoxygen affinity (Storz et al , 2010aNatarajan et al 2011), which enhances pulmonary oxygen loading under hypoxia. Thus, the deer mouse b-globin polymorphism represents a rare case in which the spatial patterning of allele frequency variation across an environmental gradient can be related to adaptive functional properties of the encoded protein.…”
Section: Discussionmentioning
confidence: 99%
“…The twolocus HBB haplotype that increases in frequency as a function of altitude (d 1 d 1 ) is associated with increased hemoglobinoxygen affinity (Storz et al , 2010aNatarajan et al 2011), which enhances pulmonary oxygen loading under hypoxia. Thus, the deer mouse b-globin polymorphism represents a rare case in which the spatial patterning of allele frequency variation across an environmental gradient can be related to adaptive functional properties of the encoded protein.…”
Section: Discussionmentioning
confidence: 99%
“…To produce rHbs for the protein engineering experiments, the α A -and β A -globin genes of A. melanogenys were synthesized by Genscript after optimizing nucleotide sequences with respect to Escherichia coli codon preferences. Gene cassettes for the α A -and β A -globin genes and the methionine aminopeptidase (MAP) gene were tandemly cloned into the custom pGM expression plasmid described by Natarajan et al (21). All rHbs were expressed in the JM109 (DE3) E. coli strain.…”
Section: Methodsmentioning
confidence: 99%
“…Nevertheless, by changing expression conditions most globins can be expressed and purified as soluble proteins [25].…”
Section: Recombinant Expression In Escherichia Colimentioning
confidence: 99%