A full-genomic sequence-verified protein-coding gene collection for Francisella tularensis

Tal Murthy; Andreas Rolfs; Yanhui Hu; Zhenwei Shi; Jacob Raphael; Donna Moreira; Fontina Kelley; Seamus McCarron; Daniel Jepson; Elena Taycher; Dongmei Zuo; Stephanie E. Mohr; Mauricio Fernandez; Leonardo Brizuela; Joshua LaBaer

doi:10.1371/journal.pone.0000577

A full-genomic sequence-verified protein-coding gene collection for Francisella tularensis

Tal Murthy, Andreas Rolfs, Yanhui Hu, Zhenwei Shi, Jacob Raphael, Donna Moreira, Fontina Kelley, Seamus McCarron, Daniel Jepson, Elena Taycher, Dongmei Zuo, Stephanie E. Mohr, Mauricio Fernandez, Leonardo Brizuela, Joshua LaBaer

Research output: Contribution to journal › Article › peer-review

21 Scopus citations

Abstract

The rapid development of new technologies for the high throughput (HT) study of proteins has increased the demand for comprehensive plasmid clone resources that support protein expression. These clones must be full-length, sequence-verified and in a flexible format. The generation of these resources requires automated pipelines supported by software management systems. Although the availability of clone resources is growing, current collections are either not complete or not fully sequence-verified. We report an automated pipeline, supported by several softwate applications that enabled the construction of the first comprehensive sequence-verified plasmid clone resource for more than 96% of protein coding sequences of the genome of F. tularensis, a highly virulent human pathogen and the causative agent of tularemia. This done resource was applied to a HT protein purification pipeline successfully producing recombinant proteins for 72% of the genes. These methods and resources represent significant technological steps towards exploiting the genomic information of F. tularensis in discovery applications.

Original language	English (US)
Article number	e577
Journal	PloS one
Volume	2
Issue number	6
DOIs	https://doi.org/10.1371/journal.pone.0000577
State	Published - Jun 27 2007
Externally published	Yes

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title = "A full-genomic sequence-verified protein-coding gene collection for Francisella tularensis",

abstract = "The rapid development of new technologies for the high throughput (HT) study of proteins has increased the demand for comprehensive plasmid clone resources that support protein expression. These clones must be full-length, sequence-verified and in a flexible format. The generation of these resources requires automated pipelines supported by software management systems. Although the availability of clone resources is growing, current collections are either not complete or not fully sequence-verified. We report an automated pipeline, supported by several softwate applications that enabled the construction of the first comprehensive sequence-verified plasmid clone resource for more than 96% of protein coding sequences of the genome of F. tularensis, a highly virulent human pathogen and the causative agent of tularemia. This done resource was applied to a HT protein purification pipeline successfully producing recombinant proteins for 72% of the genes. These methods and resources represent significant technological steps towards exploiting the genomic information of F. tularensis in discovery applications.",

author = "Tal Murthy and Andreas Rolfs and Yanhui Hu and Zhenwei Shi and Jacob Raphael and Donna Moreira and Fontina Kelley and Seamus McCarron and Daniel Jepson and Elena Taycher and Dongmei Zuo and Mohr, {Stephanie E.} and Mauricio Fernandez and Leonardo Brizuela and Joshua LaBaer",

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AU - Rolfs, Andreas

AU - Hu, Yanhui

AU - Shi, Zhenwei

AU - Raphael, Jacob

AU - Moreira, Donna

AU - Kelley, Fontina

AU - McCarron, Seamus

AU - Jepson, Daniel

AU - Taycher, Elena

AU - Zuo, Dongmei

AU - Mohr, Stephanie E.

AU - Fernandez, Mauricio

AU - Brizuela, Leonardo

AU - LaBaer, Joshua

PY - 2007/6/27

Y1 - 2007/6/27

N2 - The rapid development of new technologies for the high throughput (HT) study of proteins has increased the demand for comprehensive plasmid clone resources that support protein expression. These clones must be full-length, sequence-verified and in a flexible format. The generation of these resources requires automated pipelines supported by software management systems. Although the availability of clone resources is growing, current collections are either not complete or not fully sequence-verified. We report an automated pipeline, supported by several softwate applications that enabled the construction of the first comprehensive sequence-verified plasmid clone resource for more than 96% of protein coding sequences of the genome of F. tularensis, a highly virulent human pathogen and the causative agent of tularemia. This done resource was applied to a HT protein purification pipeline successfully producing recombinant proteins for 72% of the genes. These methods and resources represent significant technological steps towards exploiting the genomic information of F. tularensis in discovery applications.

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A full-genomic sequence-verified protein-coding gene collection for Francisella tularensis

Abstract

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