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Silent mutations at the 5′-end of the cDNA of actinoporins from the sea anemone Stichodactyla helianthus allow their heterologous overproduction in Escherichia coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
25541 43580 2007 11 PDF Available
Title
Silent mutations at the 5′-end of the cDNA of actinoporins from the sea anemone Stichodactyla helianthus allow their heterologous overproduction in Escherichia coli
Abstract

Wild-type actinoporins StnI and StnII from the sea anemone Stichodactyla helianthus, as well as their NH2-terminal six-His tagged versions, have been overproduced in Escherichia coli. Overproduction of both wild-type proteins was only possible after introducing silent mutations within the 5′-end of their original cDNA sequences. These mutations would prevent the formation of RNA secondary structures blocking the ribosome-binding site and the initiation codon. The four recombinant proteins were purified to homogeneity in milligrams amount and characterized from spectroscopic and functional points of view. All the isolated proteins behaved as the corresponding natural ones although the six-His tagged variants exhibited a decreased lytic activity. The strategy described will be useful to allow the production of mutant variants of these proteins and probably of other actinoporins.

Keywords
ANTS, 8-aminonaphthalene-1,3,6-trisulfonic acid; Ch, cholesterol; DPX, N,N′-p-xylenebispyridinium bromide; Eqt, equinatoxin; Hmg, magnificalysin; PC, phosphatidylcholine; RBS, ribosome binding site; StnI and StnII, sticholysin I and II, respectively; 6HSt
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Silent mutations at the 5′-end of the cDNA of actinoporins from the sea anemone Stichodactyla helianthus allow their heterologous overproduction in Escherichia coli
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 127, Issue 2, 1 January 2007, Pages 211–221
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