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Engineering of genetic control tools in Synechocystis sp. PCC 6803 using rational design techniques

Paper ID Volume ID Publish Year Pages File Format Full-Text
22773 43384 2015 11 PDF Available
Title
Engineering of genetic control tools in Synechocystis sp. PCC 6803 using rational design techniques
Abstract

•Leveraged rational design strategies to manipulate promoters in Synechocystis.•Developed promoters capable of a 78X range in expression in Synechocystis.•We demonstrate near full repression of an inducible promoter in Synechocystis.•Developed an inducible promoter capable of high levels of induction in Synechocystis.

Cyanobacteria show promise as photosynthetic microbial factories capable of harnessing sunlight and CO2 to produce valuable end products, but few genetic control tools have been characterized and utilized in these organisms. To develop a suite of control elements capable of gene control at a variety of expression strengths, a library of 10 promoter-constructs were developed and built via rational design techniques by adding individual nucleotides in a step-wise manner within the −10 and −35 cis-acting regions of the tac promoter. This suite produced a dynamic range of expression strength, exhibiting a 78 fold change between the lowest expressing promoter, Psca8- and the highest expressing promoter, Psca3-2 when tested within Synechocystis sp. PCC 6803. Additionally, this study details the construction of a chemically inducible construct for use in Synechocystis that is based on the tac repressor system most commonly used in Escherichia coli. This research demonstrates the construction of a highly expressed inducible promoter that is also capable of high levels of gene repression. Upon chemical induction with IPTG, this same mutant strain was capable of exhibiting an average 24X increase in GFP expression over that of the repressed state.

Keywords
Cyanobacteria; Metabolic engineering; Metabolic tool development; Synthetic biology; Rational design
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Engineering of genetic control tools in Synechocystis sp. PCC 6803 using rational design techniques
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 216, 20 December 2015, Pages 36–46
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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