A Full-Genomic Sequence-Verified Protein-Coding Gene Collection for Francisella tularensis

被引：20

作者：

Murthy, Tal ^{[1
]}

Rolfs, Andreas ^{[1
]}

Hu, Yanhui ^{[1
]}

Shi, Zhenwei ^{[1
]}

Raphael, Jacob ^{[1
]}

Moreira, Donna ^{[1
]}

Kelley, Fontina ^{[1
]}

McCarron, Seamus ^{[1
]}

Jepson, Daniel ^{[1
]}

Taycher, Elena ^{[1
]}

Zuo, Dongmei ^{[1
]}

Mohr, Stephanie E. ^{[1
,2
]}

Fernandez, Mauricio ^{[1
]}

Brizuela, Leonardo ^{[1
]}

LaBaer, Joshua ^{[1
,2
]}

机构：

[1] Harvard Univ, Sch Med, Harvard Inst Prote, Dept Biol Chem & Mol Pharmacol, Cambridge, MA 02138 USA

[2] Harvard Univ, Sch Med, DF HCC DNA Resource Core, Cambridge, MA 02138 USA

来源：

PLOS ONE | 2007年 / 2卷 / 06期

关键词：

D O I：

10.1371/journal.pone.0000577

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：7

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