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 journalArticle

18 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 languageEnglish (US)
Article numbere577
JournalPloS one
Volume2
Issue number6
DOIs
StatePublished - Jun 27 2007

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    Murthy, T., Rolfs, A., Hu, Y., Shi, Z., Raphael, J., Moreira, D., Kelley, F., McCarron, S., Jepson, D., Taycher, E., Zuo, D., Mohr, S. E., Fernandez, M., Brizuela, L., & LaBaer, J. (2007). A full-genomic sequence-verified protein-coding gene collection for Francisella tularensis. PloS one, 2(6), [e577]. https://doi.org/10.1371/journal.pone.0000577