An automated high-throughput screening method for the identification of high-yield, soluble protein variants using cell-free expression and systematic truncation.

An automated high-throughput screening method for the identification of high-yield, soluble protein variants using cell-free expression and systematic truncation. Research Abstract Details 

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An automated high-throughput screening method for the identification of high-yield, soluble protein variants using cell-free expression and systematic truncation. Abstract Text:

Evan H Bursey,Chang-Yub Kim,Minmin Yu,Thomas C Terwilliger,Li-Wei Hung,

A highly automated method for rapidly identifying soluble protein variants with good expression yields has been developed. This method is based on a commercially available in vitro protein expression system. It consists of two polymerase chain reactions (PCR) followed by in vitro protein expression and protein quantification by dot blot. The PCR protocols have been improved and optimized to allow automation using commercial fluid handling devices. A PCR primer design program has also been implemented to streamline protein variant design. This automated protocol is highly reliable and has tremendously improved the throughput of expression screening as compared to conventional cell-based methods and manual in vitro methods. We have applied this method to 32 problematic targets from the TB Structural Genomics Consortium. Experimental results of these studies are reported.

An automated high-throughput screening method for the identification of high-yield, soluble protein variants using cell-free expression and systematic truncation. Publishing Authors By Initials

EH Bursey,CY Kim,M Yu,TC Terwilliger,LW Hung,

For similar proteins: recombinant proteins research abstracts see: proteins: recombinant proteins research

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An automated high-throughput screening method for the identification of high-yield, soluble protein variants using cell-free expression and systematic truncation. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Journal of structural and functional genomics

VOLUME: 7

Page Numbers: 139-47

Journal Abbreviation: J. Struct. Funct. Genomics

ISSN: 1345-711X

DAY: 31

MONTH: 05

YEAR: 2007

An automated high-throughput screening method for the identification of high-yield, soluble protein variants using cell-free expression and systematic truncation. Information

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LANGUAGE: eng

NlmUniqueID: 101128185

An automated high-throughput screening method for the identification of high-yield, soluble protein variants using cell-free expression and systematic truncation. Keywords Mesh Terms:

KEYWORDS: Recombinant Proteins

MESH TERMS: genetics

Chemical & Substance for Abstract: An automated high-throughput screening method for the identification of high-yield, soluble protein variants using cell-free expression and systematic truncation. Information

Substance Name: Recombinant Proteins

Registry Number: 0

Grant and Affiliation Information for An automated high-throughput screening method for the identification of high-yield, soluble protein variants using cell-free expression and systematic truncation.

AFFILIATION: Physical Biosciences Division, Lawrence Berkeley National Laboratory, MS4R0230, Berkeley, CA 94720, USA.

Country: Netherlands

AGENCY: United States NIGMS

GRANT: P50 GM62410

ACRONYM: GM

MEDLINETA: J Struct Funct Genomics

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