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Development of transgenic mice harboring ovine beta lactoglobulin-calcitonin transgene
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A. R. نیاورانی   , S. دهقانیزاده , M. کریمی , S. زینلی |
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Abstract: (14626 Views) |
| Introduction: Expression of foreign proteins in mammalian milk is becoming a widespread strategy for high-level production of recombinant pharmaceuticals, especially those with the most complex post-translational modifications. We have attempted in this project to develop transgenic mice harboring a transgene driving mammary gland expression of hybrid human salmon calcitonin.
Materials and Methods: A milk-specific ovine beta-lactoglobulin (oBLG) promoter was used to drive expression of recombinant calcitonin in mouse milk. A gene construct was generated, consisting of 10.7kbp of the oBLG gene including its promoter and 3' flanking region with the calcitonin coding sequences inserted in-frame into the oBLG fifth exon. The gene construct was purified using CsCl gradient, released from vector, and gel-purified. After appropriate dilution, it was microinjected into recently-fertilized mouse oocytes. These oocytes then were transferred to pseudo-pregnant foster mice.
Results: Forty one pups were born from foster mice, which were genotyped using PCR, slot blotting, and Southern blotting. Among 9 mice which showed positive PCR results, 6 mice resulted in pups with positive PCR tests. All six families transmitted the transgene to first and second generation.
Conclusion: As the main criteria for considering a mouse as transgenic is transgene transmission to the next generation, all 6 mice which stably transmitted their transgene to progeny are considered as transgenic founders and constitute independent transgenic lines. |
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| Keywords: Transgenic mice, Milk, Calcitonin, Beta-lactoglobulin, Recombinant protein, Fusion protein |
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Full-Text [PDF 350 kb]
(3746 Downloads)
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Type of Study: Applicable |
Subject:
General
Received: 2008/09/10 | Published: 2005/05/15
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