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Title: High Throughput Methods To Identify Underlying Molecular Signature Pathways In Desulfovibrio vulgaris.

Abstract

Abstract not provided.

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Sandia National Lab. (SNL-CA), Livermore, CA (United States)
Sponsoring Org.:
USDOE National Nuclear Security Administration (NNSA)
OSTI Identifier:
1147817
Report Number(s):
SAND2007-3051C
523323
DOE Contract Number:
AC04-94AL85000
Resource Type:
Conference
Resource Relation:
Conference: Proposed for presentation at the ASM Conference held May 21-25, 2007 in Toronto, Canada.
Country of Publication:
United States
Language:
English

Citation Formats

Gaucher, Sara P, Hadi, Masood, Singh, Anup K., Light, Yooli Kim, Hazen, Terry, and ARKIN, ADAM. High Throughput Methods To Identify Underlying Molecular Signature Pathways In Desulfovibrio vulgaris.. United States: N. p., 2007. Web.
Gaucher, Sara P, Hadi, Masood, Singh, Anup K., Light, Yooli Kim, Hazen, Terry, & ARKIN, ADAM. High Throughput Methods To Identify Underlying Molecular Signature Pathways In Desulfovibrio vulgaris.. United States.
Gaucher, Sara P, Hadi, Masood, Singh, Anup K., Light, Yooli Kim, Hazen, Terry, and ARKIN, ADAM. Tue . "High Throughput Methods To Identify Underlying Molecular Signature Pathways In Desulfovibrio vulgaris.". United States. doi:. https://www.osti.gov/servlets/purl/1147817.
@article{osti_1147817,
title = {High Throughput Methods To Identify Underlying Molecular Signature Pathways In Desulfovibrio vulgaris.},
author = {Gaucher, Sara P and Hadi, Masood and Singh, Anup K. and Light, Yooli Kim and Hazen, Terry and ARKIN, ADAM},
abstractNote = {Abstract not provided.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue May 01 00:00:00 EDT 2007},
month = {Tue May 01 00:00:00 EDT 2007}
}

Conference:
Other availability
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  • Abstract not provided.
  • Our scheme for the tagless purification of water soluble complexes. 10 g of protein from a crude bacterial extract is first fractionated by ammonium sulfate precipitation and then by a series of chromatographic steps: anion exchange (IEX), hydrophobic interaction (HIC), and finally size exclusion (Gel Filtration). Fractions from the last chromatography step are trypsin digested and peptides labeled with iTRAQ reagents to allow multiplexing and quantitation during mass spectrometric analysis. Elution profiles of identified proteins are then subjected to clustering analysis.
  • The ability to conduct advanced functional genomic studies of the thousands of 38 sequenced bacteria has been hampered by the lack of available tools for making high39 throughput chromosomal manipulations in a systematic manner that can be applied across 40 diverse species. In this work, we highlight the use of synthetic biological tools to 41 assemble custom suicide vectors with reusable and interchangeable DNA parts to 42 facilitate chromosomal modification at designated loci. These constructs enable an array 43 of downstream applications including gene replacement and creation of gene fusions with 44 affinity purification or localization tags. We employed thismore » approach to engineer 45 chromosomal modifications in a bacterium that has previously proven difficult to 46 manipulate genetically, Desulfovibrio vulgaris Hildenborough, to generate a library of 47 662 strains. Furthermore, we demonstrate how these modifications can be used for 48 examining metabolic pathways, protein-protein interactions, and protein localization. The 49 ubiquity of suicide constructs in gene replacement throughout biology suggests that this 50 approach can be applied to engineer a broad range of species for a diverse array of 51 systems biological applications and is amenable to high-throughput implementation.« less
  • The ability to conduct advanced functional genomic studies of the thousands of sequenced bacteria has been hampered by the lack of available tools for making high- throughput chromosomal manipulations in a systematic manner that can be applied across diverse species. In this work, we highlight the use of synthetic biological tools to assemble custom suicide vectors with reusable and interchangeable DNA “parts” to facilitate chromosomal modification at designated loci. These constructs enable an array of downstream applications including gene replacement and creation of gene fusions with affinity purification or localization tags. We employed this approach to engineer chromosomal modifications inmore » a bacterium that has previously proven difficult to manipulate genetically, Desulfovibrio vulgaris Hildenborough, to generate a library of over 700 strains. Furthermore, we demonstrate how these modifications can be used for examining metabolic pathways, protein-protein interactions, and protein localization. The ubiquity of suicide constructs in gene replacement throughout biology suggests that this approach can be applied to engineer a broad range of species for a diverse array of systems biological applications and is amenable to high-throughput implementation.« less
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