Development of transgenic mice expressing calcitonin as a beta-lactoglobulin fusion protein in mammary gland.
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 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 10.7 kbp of the oBLG gene including its promoter and 3' flanking region with the calcitonin coding sequences inserted in-frame into the oBLG fifth exon. After microinjection, six founder mice transmitted the transgene to their progeny. RT-PCR confirmed mammary-gland specific expression of recombinant mRNA in most transgenic mice and Western blot analysis confirmed expression of chimeric protein. Calcitonin can thus be expressed under the oBLG promoter and regulatory elements in a mammary-gland specific manner.
Niavarani
Ahmadreza
A
Department of Biotechnology, Pasteur Institute of Iran, Tehran, Iran.
Dehghanizadeh
Somaye
S
Zeinali
Sirous
S
Karimi
Mohsen
M
Magliano
Marc
M
Rassoulzadegan
Minoo
M
D000818:Animals; D001483:Base Sequence; D002116:Calcitonin / Q000235:genetics* / Q000378:metabolism; D004274:DNA, Recombinant / Q000235:genetics; D005260:Female; D015870:Gene Expression; D007782:Lactoglobulins / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008321:Mammary Glands, Animal / Q000378:metabolism*; D051379:Mice; D008811:Mice, Inbred DBA; D008822:Mice, Transgenic; D008892:Milk / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012756:Sheep
D004274:DNA, Recombinant; D007782:Lactoglobulins; D011993:Recombinant Fusion Proteins; D002116:Calcitonin