fulltext.study @t Gmail

Production of Rous sarcoma virus-like particles displaying human transmembrane protein in silkworm larvae and its application to ligand–receptor binding assay

Paper ID Volume ID Publish Year Pages File Format Full-Text
23718 43467 2011 8 PDF Available
Title
Production of Rous sarcoma virus-like particles displaying human transmembrane protein in silkworm larvae and its application to ligand–receptor binding assay
Abstract

Two types of Rous sarcoma virus (RSV) group-antigen protein (Gag) virus like particles (VLPs), full-length Gag (Gag701) and RSV protease domain (PR)-deleted mutant (Gag577) were expressed in silkworm larvae. Gag577 was secreted into hemolymph efficiently using wild type bacmid (WT), cysteine protease-deficient bacmid (CP−), cysteine protease and chitinase-deficient bacmid (CP−Chi−) bacmids, but comparatively Gag701 secretion levels were low. VLPs were purified on 10–60% (v/v) sucrose density gradient by ultracentrifugation and their structures confirmed under electron microscope. When hPRR and RSV Gag577 were co-expressed in silkworm larvae, human prorenin receptor (hPRR) was displayed on the surface of RSV VLPs, which was detected by Western blotting and immunoelectron microscopy. Moreover, binding of hPRR localized on the surface of VLPs to human prorenin was confirmed by ELISA. These results indicate that active hPRR was displayed on the surface of RSV VLPs, which can be utilized for drug discovery of hPRR blockers to prevent nephropathy. Moreover, this transmembrane protein display system using RSV Gag in silkworm larvae is applicable to expression of intact transmembrane proteins and binding assay of transmembrane proteins to its ligands, especially for transmembrane proteins which cannot be purified from membrane fractions in active states.

Keywords
Rous sarcoma virus; Gag; Virus-like-particles; Surface display; Human prorenin receptor; Silkworm
First Page Preview
Production of Rous sarcoma virus-like particles displaying human transmembrane protein in silkworm larvae and its application to ligand–receptor binding assay
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 155, Issue 2, 10 September 2011, Pages 185–192
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