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A rapid protein expression and purification system using Chinese hamster ovary cells expressing retrovirus receptor

Paper ID Volume ID Publish Year Pages File Format Full-Text
25617 43585 2006 12 PDF Available
Title
A rapid protein expression and purification system using Chinese hamster ovary cells expressing retrovirus receptor
Abstract

Conventional stable protein expression systems using mammalian cells include a time-consuming step of antibiotic resistance-based cell cloning. Here, we report a rapid flow cytometry-based method for the collection of retrovirus vector-infected Chinese hamster ovary (CHO) cells that express desired proteins. The vector carries the genes for green fluorescent protein (GFP), as a marker, and glutathione-S-transferase (GST), to express the desired protein as a GST-fusion construct. To render CHO cells susceptible to retrovirus infection, they were forced to express EcoR, the receptor for retroviruses. After infection, cells expressing desired proteins were collected by flow cytometry as a GFP-positive population, and the desired proteins were purified by glutathione affinity chromatography. This method reduces the time required between infection of cells and purification of a desired protein from several months to approximately 2 weeks.

Keywords
CHO, Chinese hamster ovary; EcoR, ecotropic retrovirus receptor; GFP, green fluorescent protein; GST, glutathione-S-tarnsferase; IB, immunoblot(ting); PAGE, polyacrylamide gel electrophoresis; PCR, polymerase chain reactionCHO cells; Ecotropic retrovirus
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A rapid protein expression and purification system using Chinese hamster ovary cells expressing retrovirus receptor
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 126, Issue 4, 1 December 2006, Pages 463–474
Authors
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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
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Full-text PDF Download
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Any Questions? feel free to contact us