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Russian Journal of Genetics

, Volume 41, Issue 10, pp 1088–1094 | Cite as

Secretion of Biologically Active Human Granulocyte Colony-Stimulating Factor (G-CSF) in Milk of Transgenic Mice

  • G. A. Dvoryanchikov
  • I. A. Serova
  • L. E. Andreeva
  • L. P. B. Dias
  • S. Azevedo
  • O. L. Serov
Molecular Genetics

Abstract

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 (CSN1S1). 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 µg/ml to over 1 mg/ml, in mice with construct pGCm1 and was low (up to 60 µg/ml) or absent in mice with construct pGCm2. G-CSF glycosylation was incomplete in mice with transgene pGCm1 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.

Keywords

Transgenic Mouse Flank Region Promoter Sequence Flank Sequence Lactate Female 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© MAIK "Nauka/Interperiodica" 2005

Authors and Affiliations

  • G. A. Dvoryanchikov
    • 1
  • I. A. Serova
    • 1
    • 2
  • L. E. Andreeva
    • 3
  • L. P. B. Dias
    • 1
  • S. Azevedo
    • 4
  • O. L. Serov
    • 2
  1. 1.Institute of Biophysics, Carlos Chagas FilhoFederal University of Rio de JaneiroRio de JaneiroBrazil
  2. 2.Institute of Cytology and GeneticsRussian Academy of SciencesNovosibirskRussia
  3. 3.Institute of Molecular GeneticsRussian Academy of SciencesMoscowRussia
  4. 4.Cellular BankUniversity Hospital Clemento Fraga Filho, Federal University of Rio de JaneiroRio de JaneiroBrazil

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