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

Secretion of Biologically Active Human Granulocyte Colony-Stimulating Factor (G-CSF) in Milk of... 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Genetics Springer Journals

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

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Publisher
Nauka/Interperiodica
Copyright
Copyright © 2005 by MAIK "Nauka/Interperiodica"
Subject
Biomedicine; Microbial Genetics and Genomics; Animal Genetics and Genomics; Human Genetics
ISSN
1022-7954
eISSN
1608-3369
D.O.I.
10.1007/s11177-005-0204-8
Publisher site
See Article on Publisher Site

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.

Journal

Russian Journal of GeneticsSpringer Journals

Published: Nov 11, 2005

References

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