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Construction of a host-independent T7 expression system with small RNA regulation

Paper ID Volume ID Publish Year Pages File Format Full-Text
23063 43415 2014 4 PDF Available
Title
Construction of a host-independent T7 expression system with small RNA regulation
Abstract

•We integrate T7 RNA Polymerase gene and T7 Promoter into a single plasmid.•We use small antisense RNA to regulate T7 RNA Polymerase expression on plasmid.•We construct a novel T7 expression system independent on DE3 lysogenic hosts.•We prove the expression ability of this system in 4 non-DE3 E. coli strains.•We enhance the protein expression in an industrial wild-type strain by this system.

It is desirable to build a universal and efficient protein expression system for wild-type prokaryotic strains in biotechnology industry and the outstanding T7 expression system could be a good candidate. However, the current utilization of T7 system depends on the specific DE3 lysogenic hosts, which severely limits its application in wild-type strains. In this study, a host-independent T7 expression system without relying on DE3 lysogenic hosts to provide T7 RNA Polymerase was developed. T7 RNA Polymerase gene (Gene1) and T7 Promoter were successfully integrated into a single plasmid with the regulation of proper antisense RNA to limit T7 RNA Polymerase expression at a non-lethal level. This host-independent T7 expression system realized efficient protein expression in 4 non-DE3 Escherichia coli strains and a wild-type Sinorhizobium strain TH572.

Keywords
T7 RNA Polymerase; T7 Promoter; Antisense RNA; Host-independent expression system
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Construction of a host-independent T7 expression system with small RNA regulation
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 189, 10 November 2014, Pages 72–75
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