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A high-throughput comparison of recombinant gene expression parameters for E. coli-mediated gene transfer to P388D1 macrophage cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
24841 43540 2008 6 PDF Available
Title
A high-throughput comparison of recombinant gene expression parameters for E. coli-mediated gene transfer to P388D1 macrophage cells
Abstract

Escherichia coli strain BL21(DE3) was tested as a delivery vector for gene transfer to a murine P388D1 macrophage cell line using a 96-well high-throughput assay. Five recombinant strains of E. coli were compared to identify the effect recombinant listeriolysin O (LLO) and associated gene expression parameters had on final delivery of a luciferase reporter gene. Listeriolysin O, native to Listeria monocytogenes and used here in an effort to improve final gene delivery, was expressed from plasmid and chromosomal locations under the control of constitutive Tet or inducible T7 promoters. The E. coli vectors delivered the luciferase reporter gene to the P388D1 line with success assessed by recording luciferase luminescence activity within the macrophage cells. The assay allowed rapid analysis and evaluation of each E. coli strain tested with strain BL21(DE3) harboring a chromosomal copy of the T7-driven LLO gene showing the greatest relative measure of gene delivery. Strains were separately assayed for LLO activity and exhibited a trend of maximum gene delivery between the lowest and highest recorded LLO activities.

Keywords
Bacterial vectors; Gene delivery; E. coli; High-throughput; Listeriolysin O
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A high-throughput comparison of recombinant gene expression parameters for E. coli-mediated gene transfer to P388D1 macrophage cells
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 137, Issues 1–4, 10 October 2008, Pages 59–64
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
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us