Autor: Anita Dekker, De Wit Ineke, P. David Toman, R. Berg, Gerard Platenburg, Erika Platenburg, Naomi Sakai, Frank Pieper, Karatzas Costas N, Caroline Samuel, G. Daniels
Rok vydání: 1999
Předmět:
Zdroj: Transgenic Research. 8:415-427
ISSN: 0962-8819
DOI: 10.1023/a:1008959924856
Popis: The large scale production of recombinant collagen for use in biomaterials requires an efficient expression system capable of processing a large (>400 Kd) multisubunit protein requiring post-translational modifications. To investigate whether the mammary gland of transgenic animals fulfills these requirements, transgenic mice were generated containing the αS1-casein mammary gland-specific promoter operatively linked to 37 Kb of the human α1(I) procollagen structural gene and 3′ flanking region. The frequency of transgenic lines established was 12%. High levels of soluble triple helical homotrimeric [(α1)3] type I procollagen were detected (up to 8 mg/ml) exclusively in the milk of six out of 9 lines of lactating transgenic mice. The transgene-derived human procollagen chains underwent efficient assembly into a triple helical structure. Although proline or lysine hydroxylation has never been described for any milk protein, procollagen was detected with these post-translational modifications. The procollagen was stable in mil; minimal degradation was observed. These results show that the mammary gland is capable of expressing a large procollagen gene construct, efficiently assembling the individual polypeptide chains into a stable triple helix, and secreting the intact molecule into the milk.
Databáze: OpenAIRE
Externí odkaz: