A full-genomic sequence-verified protein-coding gene collection for Francisella tularensis.
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.
Murthy
Tal
T
Harvard Institute of Proteomics, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Cambridge, Massachusetts, United States of America.
Rolfs
Andreas
A
Hu
Yanhui
Y
Shi
Zhenwei
Z
Raphael
Jacob
J
Moreira
Donna
D
Kelley
Fontina
F
McCarron
Seamus
S
Jepson
Daniel
D
Taycher
Elena
E
Zuo
Dongmei
D
Mohr
Stephanie E
SE
Fernandez
Mauricio
M
Brizuela
Leonardo
L
LaBaer
Joshua
J
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
D001426:Bacterial Proteins; D018076:DNA, Complementary