Production of Recombinant Human Type I Procollagen Homotrimer in the Mammary Gland of Transgenic Mice

Overview

Journal Transgenic Res

Specialty Molecular Biology

Date 2000 Apr 18

PMID 10767986

Citations 11

Authors

P D Toman

F Pieper

N Sakai

C Karatzas

E Platenburg

I de Wit

C Samuel

A Dekker

G A Daniels

R A Berg

G J Platenburg

Affiliations

Soon will be listed here.

Abstract

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 alpha S1-casein mammary gland-specific promoter operatively linked to 37 Kb of the human alpha 1(I) procollagen structural gene and 3' flanking region. The frequency of transgenic lines established was 12%. High levels of soluble triple helical homotrimeric [(alpha 1)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 milk; 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.

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