Generation and validation of a Shewanella oneidensis MR-1 clone set for protein expression and phage display.

PloS One
Haichun GaoJizhong Zhou

Abstract

A comprehensive gene collection for S. oneidensis was constructed using the lambda recombinase (Gateway) cloning system. A total of 3584 individual ORFs (85%) have been successfully cloned into the entry plasmids. To validate the use of the clone set, three sets of ORFs were examined within three different destination vectors constructed in this study. Success rates for heterologous protein expression of S. oneidensis His- or His/GST-tagged proteins in E. coli were approximately 70%. The ArcA and NarP transcription factor proteins were tested in an in vitro binding assay to demonstrate that functional proteins can be successfully produced using the clone set. Further functional validation of the clone set was obtained from phage display experiments in which a phage encoding thioredoxin was successfully isolated from a pool of 80 different clones after three rounds of biopanning using immobilized anti-thioredoxin antibody as a target. This clone set complements existing genomic (e.g., whole-genome microarray) and other proteomic tools (e.g., mass spectrometry-based proteomic analysis), and facilitates a wide variety of integrated studies, including protein expression, purification, and functional analyses of proteins both in viv...Continue Reading

References

Nov 7, 1995·Gene·K S Reece, G J Phillips
Apr 4, 2000·Current Opinion in Microbiology·J A Hoch
Nov 15, 2000·Genome Research·J L HartleyM A Brasch
Aug 30, 2002·Applied and Environmental Microbiology·Anton A KorenevskyTerry J Beveridge
Oct 9, 2002·Nature Biotechnology·John F HeidelbergClaire M Fraser
Sep 26, 2003·Omics : a Journal of Integrative Biology·N DaraseliaT Tatusova
Sep 27, 2003·Analytical Biochemistry·Bahram AreziHolly H Hogrefe
Jan 6, 2004·Science·Siming LiMarc Vidal
Oct 19, 2004·Genome Research·Gerald Marsischky, Joshua LaBaer
Oct 19, 2004·Genome Research·João Carlos AguiarDaniel J Carucci
Oct 19, 2004·Genome Research·Joshua LabaerLeonardo Brizuela
Oct 19, 2004·Genome Research·Amélie DricotXavier De Bolle
Nov 2, 2004·Journal of Bacteriology·Haichun GaoJizhong Zhou
Jan 27, 2005·Omics : a Journal of Integrative Biology·Margaret F RomineMary S Lipton
Feb 3, 2005·Proceedings of the National Academy of Sciences of the United States of America·Eugene KolkerJames M Tiedje
Feb 26, 2005·Nature Genetics·Mamie Z Li, Stephen J Elledge
Mar 19, 2005·Journal of Bacteriology·Yongqing LiuJizhong Zhou
May 11, 2005·Molecular Microbiology·Jeffrey A GralnickDianne K Newman
Jun 2, 2005·Proceedings of the National Academy of Sciences of the United States of America·Jaehong ParkLeonardo Brizuela
Jun 24, 2005·Infection and Immunity·Matthew McKevittTimothy Palzkill
Nov 5, 2005·Applied and Environmental Microbiology·Rizlan Bencheikh-LatmaniBradley M Tebo
Nov 10, 2005·Chemical Reviews·John W Kehoe, Brian K Kay
Dec 3, 2005·Genes & Development·Daniel M GelperinElizabeth J Grayhack
Jan 5, 2006·Applied and Environmental Microbiology·Steven D BrownDorothea K Thompson
Mar 10, 2006·Molecular & Cellular Proteomics : MCP·Steven D BrownDorothea K Thompson
Aug 17, 2006·Critical Reviews in Biochemistry and Molecular Biology·Eric M Phizicky, Elizabeth J Grayhack
Sep 8, 2006·Applied and Environmental Microbiology·Karuna ChoureyDorothea K Thompson
Sep 20, 2006·Molecular & Cellular Proteomics : MCP·Nazila Salamat-MillerC Russell Middaugh
Nov 14, 2006·Applied and Environmental Microbiology·Yinjie J TangJay D Keasling

❮ Previous
Next ❯

Citations

Dec 3, 2014·Protein Engineering, Design & Selection : PEDS·Soili I LehtonenVesa P Hytönen
Apr 2, 2013·Journal of Bacteriology·Linlin SunHaichun Gao
Feb 19, 2013·Environmental Microbiology·Huihui FuHaichun Gao

❮ Previous
Next ❯

Datasets Mentioned

BETA
AE014300

Methods Mentioned

BETA
nuclear magnetic resonance
PCR
electrophoretic mobility
phage display
phage
two-hybrid
ELISA
transfections

Software Mentioned

BLASTx
BLASTn
PRIMEGENS
TIGR
MAGIC

Related Concepts

Related Feeds

Aminoglycosides (ASM)

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.

Bacterial Protein Structures

Bacterial protein structures can expedite the development of novel antibiotics. Here is the latest research on bacterial proteins and the resolution of their structures.

Aminoglycosides

Aminoglycoside is a medicinal and bacteriologic category of traditional Gram-negative antibacterial medications that inhibit protein synthesis and contain as a portion of the molecule an amino-modified glycoside. Discover the latest research on aminoglycoside here.

Brucellosis (ASM)

Brucellosis is a bacterial infection caused by members of the genus brucella and remains one of the world's major zoonotic diseases. Discover the latest research on Brucellosis here.

Bacteriophage: Phage Therapy

Phage therapy uses bacterial viruses (bacteriophages) to treat bacterial infections and is widely being recognized as an alternative to antibiotics. Here is the latest research.

Brucellosis

Brucellosis is a bacterial infection caused by members of the genus brucella and remains one of the world's major zoonotic diseases. Discover the latest research on Brucellosis here.