Expression of functional recombinant human factor IX in milk of mice.
Human factor IX is synthesized in the liver and secreted in the blood, where it participates in a group of reactions involving coagulation factors and proteins that permit sanguinary coagulation. In this work two lines of transgenic mice were developed to express the FIX gene in the mammalian glands under control of milk beta-casein promoter. The founding females secreted the FIX in their milk (3% total soluble protein). The stable integration of transgene was confirmed by southern blot analysis. The presence of the FIX recombinant protein in the milk of transgenic females was confirmed by western blot and the clotting activity was revealed in blood-clotting assays. The coagulation activity in human blood treated with recombinant FIX increased while the time of coagulation decreased. Our results confirm the production of a large amount of recombinant biologically active FIX in the mammary gland of transgenic mice.
Lisauskas
Sharon F C
SF
Embrapa Recursos Genéticos e Biotecnologia, Parque Estação Biológica, PqEB, Av. W5 Norte (final), Brasilia, DF, CEP 70770-900, Brazil.
Cunha
Nicolau B
NB
Vianna
Giovanni R
GR
Mendes
Erica A
EA
Ramos
Gustavo L
GL
Maranhão
Andréia Q
AQ
Brígido
Marcelo M
MM
Almeida
Jussara O S C
JO
Baptista
Heloisa A
HA
Motta
Fabiana L T
FL
Pesquero
João B
JB
Aragão
Francisco J L
FJ
Rech
Elíbio L
EL
D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D005164:Factor IX / Q000096:biosynthesis* / Q000378:metabolism / Q000502:physiology; D005260:Female; D007774:Lactation; D008297:Male; D008321:Mammary Glands, Animal / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008894:Milk Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D010314:Partial Thromboplastin Time; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism
D008894:Milk Proteins; D011994:Recombinant Proteins; D005164:Factor IX