Development of Transgenic Mice Harboring Ovine Beta Lactoglobulin-Calcitonin Transgene

Document Type: Research Paper

Authors

1 Department of Biotechnology, Pasteur Institute of Iran, Tehran, I.R. Iran.

2 INSERM U636, University of Nice-Sophia Antipolis, France.

Abstract

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.
A gene construct was generated, consisting of 10.7 kbp of the ovine beta-lactoglobulin (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. It
was microinjected into fertilized mouse oocytes. These oocytes were then transferred to pseudo-pregnant foster mice. The pups born from foster mice were genotyped using PCR, slot blot, and Southern blot techniques. Among 9 mice which showed positive PCR results, only 6 mice transmitted the transgene to the
next generation. Therefore, 6 transgenic lines were established which stably transmitted their transgene to
their progeny.

Keywords