Synthesis and characterization of biologically active recombinant elk and horse FSH.
The objective of this investigation was to clone and express the elk and horse common alpha-subunit and FSH beta-subunit cDNAs, and to produce recombinant FSH from both species in vitro. The RNAs extracted from elk and horse pituitary glands were reverse-transcribed and amplified by polymerase chain reaction. The cDNAs corresponding to both subunits of elk and horse were cloned into the expression vector pBudCE4.1 and transfected into CRL-9096 cells. Expression of both genes was determined in the transfected cells by Northern and Western blot analysis. Recombinant elk and horse FSH secreted in culture media were characterized by an in vitro bioassay and RIA. When the recombinant products were assessed as activity over mass of FSH measured by RIA, the horse product was 5.6 times more potent than the elk product. The recombinant products injected to immature female Wistar rats stimulated ovarian growth. The results suggest that the products obtained correspond to recombinant versions of the native elk and horse FSH. The availability of these recombinant products may aid in the development of more predictable and efficient techniques of ovarian stimulation in cervids, equids, and other species as well.
Fachal
María Victoria
MV
Department of Biology, University of Saskatchewan College of Art and Sciences, Saskatoon, Canada.
Furlan
Mike
M
Clark
Rena
R
Card
Claire E
CE
Chedrese
P Jorge
PJ
D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D018076:DNA, Complementary / Q000302:isolation & purification; D005640:Follicle Stimulating Hormone / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics* / Q000502:physiology; D006736:Horses / Q000235:genetics*; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D012418:Ruminants / Q000235:genetics*; D014162:Transfection
D018076:DNA, Complementary; D011994:Recombinant Proteins; D005640:Follicle Stimulating Hormone