Production of human embryonic haemoglobin (Gower II) in a yeast expression system.
The cDNA coding for a human embryonic globin protein has been obtained from an erythroleukaemic cell line. A plasmid expression system for human embryonic haemoglobin Gower II containing cDNA copies of the appropriate pair of globin genes coupled to synthetic galactose-regulated hybrid promoters has been engineered. Transformation of Saccharomyces cerevisiae with this plasmid yields a cellular system capable of high-level production of fully functional tetrameric embryonic haemoglobin. We have developed a purification scheme which gives high yields of pure human embryonic haemoglobin suitable for structural and functional studies. Preliminary characterization studies are reported.
Mould
R M
RM
Department of Biochemistry, University of Auckland, New Zealand.
Hofmann
O M
OM
Brittain
T
T
D000595:Amino Acid Sequence; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D018014:Gene Transfer Techniques; D005914:Globins / Q000737:chemistry / Q000235:genetics; D006455:Hemoglobins, Abnormal / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D010100:Oxygen / Q000378:metabolism; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D006455:Hemoglobins, Abnormal; D046911:Macromolecular Substances; D011994:Recombinant Proteins; C030446:hemoglobin Gower; D005914:Globins; D010100:Oxygen