High-level production of human alpha- and beta-globins in insect cells.
High-level production of human alpha- and beta-globins in cultured Spodoptera frugiperda (Sf-9) cells infected with recombinant baculoviruses is described. The expressed globins are produced to 70-140 mg protein/liter of cell culture or 5-10% of the total cellular protein. Two recombinant baculoviruses for alpha-globin, H alpha and H beta alpha, differ in their construction in that the 5'-untranslated region of the beta-globin gene is inserted 5' to the alpha-globin mRNA coding region in H beta alpha. This insertion results in a 40% increase in yield of alpha-globin over that of H alpha. Consistent with previous observations of the processing of recombinant proteins in Sf-9 cells, both alpha- and beta-globins expressed in Sf-9 cells are correctly processed to remove the initiating methionine from the amino termini of the globins. Sequencing of the expressed globins in Sf-9 cells confirms their identity with globins purified from human normal adult hemoglobin.
Groebe
D R
DR
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213.
Busch
M R
MR
Tsao
T Y
TY
Luh
F Y
FY
Tam
M F
MF
Chung
A E
AE
Gaskell
M
M
Liebhaber
S A
SA
Ho
C
C
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D015870:Gene Expression; D005822:Genetic Vectors; D005914:Globins / Q000096:biosynthesis* / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D009036:Moths; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D017385:Sequence Homology
D011993:Recombinant Fusion Proteins; D005914:Globins