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Evaluating regulatory elements of human cytomegalovirus major immediate early gene for enhancing transgene expression levels in CHO K1 and HEK293 cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
24159 43502 2010 4 PDF Available
Title
Evaluating regulatory elements of human cytomegalovirus major immediate early gene for enhancing transgene expression levels in CHO K1 and HEK293 cells
Abstract

The upstream regulatory sequence (URS), NF1 region, enhancer, promoter, 1st exon, and intron A of human cytomegalovirus major immediate early gene (hCMV MIE) are evaluated for enhancing transient and stable gene expression levels in two industrial cell lines, CHO K1 and HEK293 using firefly luciferase (Fluc) and erythropoietin (EPO). As compared to the control vector which only contains the enhancer and promoter (EP), vectors containing the 1st exon (EPE) and intron A (EPEI) enhance transient expression levels of the two proteins by approximately 2.5- to 4.3-fold in the two cell lines. Addition of NF1 and URS to EP (NEP and UNEP) or EPEI (NEPEI and UNEPEI) results in a lesser effect on the expression. In stable transfections, UNEPEI provides the highest expression level in CHO K1 cells, yielding approximately 4.0-fold increase in Fluc expression and 2.5-fold increase in EPO expression. In HEK293 cells, EPE is the best and enhances Fluc and EPO expression by more than 2.0-fold. Such information is valuable for the development of optimal vectors to enhance transient and stable production of recombinant proteins in CHO K1 and HEK293 cells.

Keywords
hCMV MIE; 1st Exon; Intron A; HEK293; CHO K1; Erythropoietin
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Evaluating regulatory elements of human cytomegalovirus major immediate early gene for enhancing transgene expression levels in CHO K1 and HEK293 cells
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 147, Issues 3–4, June 2010, Pages 160–163
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