A high efficiency cloning and expression system for proteomic analysis.
The recent description of the complete genomes of the two most pathogenic species of Brucella opens the way for genome-based analysis of the antigenicity of their proteins. In the present report, we describe a bench-level high-efficiency cloning and expression system (HECES) that allow expression of large numbers of Brucella proteins based on genomic sequence information. Purified proteins are produced with high efficiency in a microarray format conducive to analysis of their sero-reactivity against serum from immunized animals. This method is applicable at either small or large scale of protein processing. While it does not require robotics, the format is amenable to robotic implementation for all aspects of the process and subsequent analysis of protein characteristics. This method will allow selection of new reagents for diagnosis of brucellosis and development of vaccine against Brucella, an important zoonotic disease and biothreat agent.
Ding
Xuan Z
XZ
Department of Bacterial Diseases, Division of Communicable Diseases and Immunology, Walter Reed Army Institute of Reseach, Silver Spring, MD 20910-7500, USA. tom.ding@na.amedd.army.mil
Paulsen
Ian T
IT
Bhattacharjee
Apurba K
AK
Nikolich
Mikeljon P
MP
Myers
Gary
G
Hoover
David L
DL
D001331:Automation; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D002002:Brucella / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D011994:Recombinant Proteins