Baculovirus-mediated expression of chicken growth hormone.
A full-length chicken growth hormone (cGH) cDNA was placed downstream from the Autograph californica nuclear polyhedron virus, AcNPV, polyhedron gene promoter and expressed in Sf9 insect cells. Secreted recombinant cGH levels averaged 2-10 micrograms/ml from day 5-10 postinfection. The recombinant cGH analyzed by SDS-PAGE gels and Western blotting consisted of a doublet with M(r) of 26.5 and 23.5 kDa. Analysis by 2-D electrophoresis of partially-purified recombinant cGH and purified native cGH revealed similar immunoreactive charge isoforms and M(r) variants. The recombinant hormone was biologically active in a homologous radioreceptor assay. The results show that cGH expressed in insect cells is biologically and immunologically active, and that a variety of isoforms are secreted which exhibit size and charge properties similar to those of pituitary-derived cGH.
Foster
D N
DN
Department of Animal Science, University of Minnesota, St. Paul 55108, USA.
Proudman
J A
JA
Harmon
S A
SA
Foster
L K
LK
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D002460:Cell Line; D002645:Chickens; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D013006:Growth Hormone / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D007700:Kinetics; D011869:Radioligand Assay; D011994:Recombinant Proteins; D018411:Spodoptera; D014162:Transfection
D011994:Recombinant Proteins; D013006:Growth Hormone