fulltext.study @t Gmail

Generation of hybrid transgenic silkworms that express Bombyx mori prolyl-hydroxylase α-subunits and human collagens in posterior silk glands: Production of cocoons that contained collagens with hydroxylated proline residues

Paper ID Volume ID Publish Year Pages File Format Full-Text
25669 43589 2006 15 PDF Available
Title
Generation of hybrid transgenic silkworms that express Bombyx mori prolyl-hydroxylase α-subunits and human collagens in posterior silk glands: Production of cocoons that contained collagens with hydroxylated proline residues
Abstract

Prolyl 4-hydroxylase (P4H) is a heterotetramer enzyme consisting of α-subunits (P4Hα) and β-subunits (P4Hβ), and is required for collagen biosynthesis. Previously, we generated transgenic silkworms that produced human type III collagen fragments (mini-collagens) in the posterior silk gland (PSG). However, prolyl 4-hydroxylation did not occur on the mini-collagens, because in spite of an abundant expression of P4Hβ in PSGs, P4Hα expression was quite low there, thus resulting in an insufficient activity of P4H. In this study we aimed at generating hybrid transgenic silkworms whose PSGs are capable of producing mini-collagens and enough P4H for their prolyl 4-hydroxylation. Isolated PSGs were bombarded with fibroin L-chain gene promoter-driven vectors containing Bombyx mori P4Hα (BmP4Hα) cDNAs and were transplanted into the hemolymphatic cavity. The P4H activity in the PSG cells significantly increased, indicating that the expressed BmP4Hα formed active tetramers with endogenous BmP4Hβ. Using germ-line transgenesis technology, silkworms were generated that synthesized BmP4Hα in PSG cells. The P4H activity in the transgenic silkworms was 130-fold higher than that of wild-type counterparts. Finally, we generated hybrid transgenic silkworms that expressed cDNAs of both BmP4Hα and mini-collagen in PSG cells. They spun cocoons that contained mini-collagens whose appropriate proline residues had been adequately hydroxylated.

Keywords
BmP4Hα, Bombyx mori prolyl 4-hydroxylase α-subunit; BmP4Hβ, Bombyx mori prolyl 4-hydroxylase β-subunit; PDI, protein disulfide isomerase; P4H, prolyl 4-hydroxylase; P4Hα, prolyl 4-hydroxylase α-subunit; P4Hβ, prolyl 4-hydroxylase β-subunit; PSG, posterior
First Page Preview
Generation of hybrid transgenic silkworms that express Bombyx mori prolyl-hydroxylase α-subunits and human collagens in posterior silk glands: Production of cocoons that contained collagens with hydroxylated proline residues
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 126, Issue 2, 1 November 2006, Pages 205–219
Authors
, , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us