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publicationDate 1998-08-25-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber US-5798227-A
titleOfInvention Co-expression of alpha and beta globins
abstract The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained. We further describe the production of tetrameric human hemoglobin and di-alpha/beta2 hemoglobin in the yeast Saccharomyces cerevisiae. The synthesis of the protein is directed by a synthetic promotor consisting of two functional parts, an upstream activator sequence (UAS) that confers inducible transcription by galactose from a consensus yeast transcriptional initiation site. The expression construct is designed such that translation is expected to initiate at the same position as the human wild-type genes for alpha - and beta -globin. Three different types of expression vectors have been used: (1) alpha -globin and beta -globin contained on two separate plasmids (pGS4688 and pGS4988) in a diploid yeast strain; (2) alpha -globin and beta -globin each contained on a single plasmid (pGS289 and pGS389) and expressed in either haploid or diploid strains; and (3) di-alpha-globin and beta globin contained on a single plasmid (pGS 3089)and expressed in haploid strains. Finally, we describe the co-expression of alpha and beta globin chains. The chains are folded together and combined intracellularly with heme to form active tetrameric hemoglobin. The hemoglobin may be recovered from the cells' soluble fraction. The invention thus obviates the need to express alpha and beta globin separately, solubilize, renature and purify them, and combine them in vitro with heme to obtain an artificial hemoglobin. By way of comparison, the separately expressed beta globin known in the art is deposited in inclusion bodies. Polycistronic co-expression of alpha and beta globins is particularly preferred.
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http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP17647
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP67822
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID67989504
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ8P5Z7
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02072
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID7009579
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP67821
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID394349
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID61536005
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID67989503
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID439357
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP75089
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID610255
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02070
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ82680
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP67820
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID129915704
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02066
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ82630
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP67819
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP68082
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID135942679
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02106
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP17540
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID128937230
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID419586423
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP68081
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID107371
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ2MJN4
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02095
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID129105737
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID281407
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP68080
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID425978018
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02094
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID123938
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ55863
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02093
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ95222
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID24440
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCB0B879
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02092
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCA3RM19
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ42690
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02090
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID56470774
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP17709
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02087
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ6D7C4
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ2PE78
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP46649
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP41359
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCO04879
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ8P8E9
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID128841
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ6D736
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02086
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID127788218
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID128837
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ944G9
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ1I4P8
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID128842119
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ2PE47
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02115
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID419586922
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID7408322
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02114
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID101841186
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCB7LL88
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID10353878
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID39251
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID416652
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCB5R3T6
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID426110856
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02113
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID22840584
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP41332
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP67824
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02112
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP36623
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ6G670
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ95KP3
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ82994
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID504174
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP67823
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP41328
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02111
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCA4FZL2
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID65359
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ42954
http://rdf.ncbi.nlm.nih.gov/pubchem/compound/CID656894
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID912048
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02110
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID24464
http://rdf.ncbi.nlm.nih.gov/pubchem/gene/GID3558
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP67974
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID419587403
http://rdf.ncbi.nlm.nih.gov/pubchem/substance/SID226394880
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP02108
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP46796
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCP67973
http://rdf.ncbi.nlm.nih.gov/pubchem/protein/ACCQ07637

Total number of triples: 2664.