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 software 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 clone 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.
authors.affiliation: Harvard Institute of Proteomics, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Cambridge, Massachusetts, United States of America.
authors.forename: Tal
authors.initials: T
authors.lastname: Murthy
authors.forename: Andreas
authors.initials: A
authors.lastname: Rolfs
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Hu
authors.forename: Zhenwei
authors.initials: Z
authors.lastname: Shi
authors.forename: Jacob
authors.initials: J
authors.lastname: Raphael
authors.forename: Donna
authors.initials: D
authors.lastname: Moreira
authors.forename: Fontina
authors.initials: F
authors.lastname: Kelley
authors.forename: Seamus
authors.initials: S
authors.lastname: McCarron
authors.forename: Daniel
authors.initials: D
authors.lastname: Jepson
authors.forename: Elena
authors.initials: E
authors.lastname: Taycher
authors.forename: Dongmei
authors.initials: D
authors.lastname: Zuo
authors.forename: Stephanie E
authors.initials: SE
authors.lastname: Mohr
authors.forename: Mauricio
authors.initials: M
authors.lastname: Fernandez
authors.forename: Leonardo
authors.initials: L
authors.lastname: Brizuela
authors.forename: Joshua
authors.initials: J
authors.lastname: LaBaer
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D005604:Francisella tularensis / Q000235:genetics* / Q000254:growth & development; D005796:Genes / Q000235:genetics*; D016680:Genome, Bacterial; D006801:Humans; D016366:Open Reading Frames / Q000235:genetics*; D014406:Tularemia / Q000235:genetics* / Q000382:microbiology / Q000473:pathology
chemical_list: D001426:Bacterial Proteins; D018076:DNA, Complementary