Linked Life Data

20661923

Source:http://linkedlifedata.com/resource/pubmed/id/20661923

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
2010-12-16
pubmed:abstractText
We describe a miniaturized fluid array device for high-throughput cell-free protein synthesis (CFPS), aiming to match the throughput and scale of gene discovery. Current practice of using E. coli cells for production of recombinant proteins is difficult and cost-prohibitive to implement in a high-throughput format. As more and more new genes are being identified, there is a considerable need to have high-throughput methods to produce a large number of proteins for studying structures and functions of the corresponding genes. The device consists of 96 units and each unit is for expression of one protein; thus up to 96 proteins can be produced simultaneously. The function of the fluid array was demonstrated by expression of a variety of proteins, with more than two orders of magnitude reduction in reagent consumption compared with a commercially available CFPS instrument. The protein expression yield in the device was up to 87 times higher for ?-glucoronidase than that in a conventional microplate. The concentration of ?-galactosidase expressed in the device was determined at 5.5 ?g/?L. The feasibility of using the device for drug screening was demonstrated by measuring the inhibitory effects of mock drug compounds on synthesized ?-lactamase without the need for harvesting proteins, which enabled us to reduce the analysis time from days to hours.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:issn
1520-6033
pubmed:author
pubmed:copyrightInfo
Copyright © 2010 American Institute of Chemical Engineers (AIChE).
pubmed:issnType
Electronic
pubmed:volume
26
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1590-6
pubmed:meshHeading
pubmed:articleTitle
Miniaturized fluid array for high-throughput protein expression.
pubmed:affiliation
Dept. of Biomedical Engineering, University of Florida, PO Box 116131, Gainesville, FL 32611, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, N.I.H., Extramural