[Secretion of biologically active human granulocyte colony-stimulating factor (G-CSF) in milk of transgenic mice].
Two constructs were devised, containing the full-length gene of the human granulocyte colony-stimulating factor (G-CSF) fused with the 5' and 3' flanking promoter sequences of bovine alpha-S1-casein gene. Both constructs contained a 1518-bp fragment that included exons 18 and 19 and 320 bp of the 3' flanking region of bovine gene @CSN1S1, but differed in size of the 5' flanking sequences, which were of 721 bp, and exon 1 in construct pGCm1 and 2001 bp and exon 1 and intron 1 in construct pGCm2. With both constructs, transgenic mice were produced. The transgene expression was assessed using RT-PCR and immunochemically from the production of human G-CSF in milk of lactating females. Secretion of human G-CSF into the milk varied in a wide range, from 0.8 microg/ml to over 1 mg/ml, in mice with construct pGCml and was low (up to 60 microg/ml) or absent in mice with construct pGCm2. G-CSF glycosylation was incomplete in mice with transgene pGCml and complete in mice with pGCm2. G-CSF of transgenic mouse milk was shown to stimulate the formation and growth of granulocyte-containing colonies in human umbilical blood cell culture and be close or identical in physiological activity to the natural human G-CSF.
Dvorianchikov
G A
GA
Serova
I A
IA
Andreeva
L E
LE
Dias
L P B
LP
Azevedo
S
S
Serov
O L
OL
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D005260:Female; D005312:Fetal Blood / Q000166:cytology / Q000502:physiology; D015870:Gene Expression; D006031:Glycosylation; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007774:Lactation / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D023461:Myeloid Progenitor Cells / Q000166:cytology / Q000502:physiology; D046188:Protein Modification, Translational / Q000235:genetics*; D011994:Recombinant Proteins; D019076:Transgenes / Q000502:physiology*
D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor