Production of the mouse whey acidic protein in transgenic pigs during lactation

Shamay, Avi; Solinas, Sabina; Pursel, V. G.; McKnight, Robert A.; Alexander, Lee; Beattie, Craig W.; Hennighausen, Lothar; Wall, Robert J.

doi:10.2527/1991.69114552x

Cited by 49 publications

(15 citation statements)

References 6 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Sci. USA 89 (1992) 12007 studies high-level production ofheterologous proteins in milk was achieved only in transgenic animals carrying either genomic (2,17) or minigene constructs (3). Our results show that a cDNA may be used to obtain high-level expression of heterologous proteins in the milk of transgenic pigs.…”

mentioning

confidence: 83%

“…A constant level of WAP of about 1 g/liter was found in milk of transgenic pigs over a 26-day lactation (17); rhPC (polyclonal population) increased about 2-fold during the first 26 days and >5-fold during the entire 55-day lactation ofpig 29-2, whereas the rhPC in the milk of pig 83-1 varied about 2.5-fold. Since WAP and rhPC transgenic pigs contained about 10-20 copies of the transgene, differences in regulation may have been due to the location of integration as well as the differences between the WAP and WAP-hPC gene structures.…”

mentioning

confidence: 99%

See 1 more Smart Citation

High-level expression of a heterologous protein in the milk of transgenic swine using the cDNA encoding human protein C.

Velander

Johnson

Page

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

150

View full text Add to dashboard Cite

Transgenic pigs were generated that produced human protein C in their milk at up to 1 g/liter. The gene construct was a fusion gene consisting of the cDNA for human protein C inserted into the first exon of the mouse whey acidic protein gene. These results demonstrate that the mouse whey acidic protein gene contains regulatory elements that can direct cDNA expression at high levels in the pig mammary gland. Recombinant human protein C that was produced at about 380 pg/ml per hr in transgenic pig milk possessed anticoagulant activity that was equivalent to that of protein C derived from human plasma. These studies provide evidence that y-carboxylation can occur at high levels in the mammary gland of a pig.

show abstract

mentioning

confidence: 83%

mentioning

confidence: 99%

High-level expression of a heterologous protein in the milk of transgenic swine using the cDNA encoding human protein C.

Velander

Johnson

Page

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

150

View full text Add to dashboard Cite

show abstract

“…After transfer and blocking (5% bovine serum albumin and 2% nonfat milk, 20 mM Tris-HCl, pH 7.6/137 mM NaCl) at room temperature for 1 h, the membranes were incubated with primary antibodies (1:1000 diluted) as follows. Stat5a (14), WAP (28) , Akt (Cell Signaling), and actin (1:4000, Chemicon). All primary antibodies were applied for 1 h at room temperature or at 4°C overnight.…”

Section: Methodsmentioning

confidence: 99%

Mammary Gland Remodeling Depends on gp130 Signaling through Stat3 and MAPK

Zhao

Hart

Cheng

et al. 2004

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

The interleukin-6 (IL6) family of cytokines signals through the common receptor subunit gp130, and subsequently activates Stat3, MAPK, and PI3K. Stat3 controls cell death and tissue remodeling in the mouse mammary gland during involution, which is partially induced by IL6 and LIF. However, it is not clear whether Stat3 activation is mediated solely through the gp130 pathway or also through other receptors. This question was explored in mice carrying two distinct mutations in the gp130 gene; one that resulted in the complete ablation of gp130 and one that led to the loss of Stat3 binding sites (gp130⌬/⌬). Deletion of gp130 specifically from mammary epithelium resulted in a complete loss of Stat3 activity and resistance to tissue remodeling comparable to that seen in the absence of Stat3. A less profound delay of mammary tissue remodeling was observed in gp130⌬/⌬ mice. Stat3 tyrosine and serine phosphorylation was still detected in these mice suggesting that Stat3 activation could be the result of gp130 interfacing with other receptors. Experiments in primary mammary epithelial cells and transfected COS-7 cells revealed a p44/42 MAPK and EGFR-dependent Stat3 activation. Moreover, the gp130-dependent EGFR activation was independent of EGF ligands, suggesting a cytoplasmic interaction and cross-talk between these two receptors. These experiments establish that two distinct Stat3 signaling pathways emanating from gp130 are utilized in mammary tissue.

show abstract

“…1) and p-lactoglobulin based transgenes provided insight into the dynamics of transgene expression in the mammary gland. Most interestingly, regulatory elements from the mouse WAP gene appear to drive expression much more efficiently in transgenic pigs than in transgenic mice Shamay et al, 1991;McKnight et al, 1992a1, suggesting that it may be of advantage to use cross species regulatory elements for mammary expression. From the production point of view this has been confirmed with human protein C. A hybrid gene consisting of the mouse WAP gene and a protein C cDNA inserted into the first WAP gene exon ( Fig.…”

Section: Expression Of Transgenes In Mammary Tissuementioning

confidence: 99%

The prospects for domesticating milk protein genes

Hennighausen¹

1992

J of Cellular Biochemistry

Self Cite

View full text Add to dashboard Cite

It is possible to convert milk glands of transgenic animals into bioreactors producing heterologous proteins such as scarce human pharmaceuticals. To predictably and successfully engineer the milk gland, we will need a thorough understanding of its physiology. Expression studies in transgenic animals have located mammary specific and hormone inducible transcription elements in the promoter/upstream regions of milk protein genes, and transfection studies in cell lines or primary cells have identified constitutive and hormone inducible elements. Most importantly, it appears that in addition to individual promoter based transcription elements structural features of milk protein chromosomal loci may contribute to the tight developmental and hormonal regulation. I will discuss milk protein gene regulation with emphasis on regulatory differences between genes and species, and the possibility that transcription elements function only properly within genetically defined chromatin domains. Novel strategies to build mammary expression vectors and to test their functionality without pursuing the standard transgenic route will be presented. Finally, I will discuss homologous recombination with the goal to target milk protein genes. Only through the domestication of milk protein genes will we be able to use their full potential in the mammary bioreactor.

show abstract

Production of the mouse whey acidic protein in transgenic pigs during lactation

Cited by 49 publications

References 6 publications

High-level expression of a heterologous protein in the milk of transgenic swine using the cDNA encoding human protein C.

High-level expression of a heterologous protein in the milk of transgenic swine using the cDNA encoding human protein C.

Mammary Gland Remodeling Depends on gp130 Signaling through Stat3 and MAPK

The prospects for domesticating milk protein genes

Contact Info

Product

Resources

About