U.S. flagAn official website of the United States government
                    Here's how you know **Official websites use .gov**
                            
                            A **.gov** website belongs to an official government organization in the United States.
 **Secure .gov websites use HTTPS**
                            
                            A **lock** (                            
                              A locked padlock                                                      
                            
                            ) or **https://** means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.


Menu Agricultural Research Service
						
                    U.S. DEPARTMENT OF AGRICULTURE
 Infectious Bacterial Diseases Research: Ames, IA
 close



    Research

                    Research Projects
                                    Publications
                                    Collaborations
                                    News
                People

                    Staff Listing
                                    CareersSearch small
                    Search
            
            ARS Home
                                About ARS
                                Contact Us




ARS Home Midwest Area Ames, Iowa National Animal Disease Center Infectious Bacterial Diseases Research Research Publications at this Location





- ARS National Programs
- Search for a scientific manuscript
- Search for a research project
- ARS Software
- Programs and Projects
- Collaborations
- Subjects of Investigation
- Publications

Title:   Author  item
                                            Bannantine, Johnitem
                                            Waters, Wadeitem
                                            Stabel, Judithitem
                                            Palmer, Mitchellitem
                                            LI, LINGLING - UNIV. OF MNitem
                                            KAPUR, VIVEK - UNIV. OF MNitem
                                            Paustian, Michael
  Submitted to: Proteomics
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 10/23/2007
                                    Publication  Date: 2/1/2008
                                    Citation: Bannantine, J.P., Waters, W.R., Stabel, J.R., Palmer, M.V., Li, L., Kapur, V., Paustian, M. 2008. Development and Use of a Partial Mycobacterium avium subspecies paratuberculosis Protein Array. Proteomics. 8(3):463-474.Interpretive Summary: This article reports on our very initial efforts at developing a protein array for M. paratuberculosis, the bacterium that causes Johne’s disease.  We cloned a set of 43 genes from this genome and recombinant produced protein from each of these clones.  The article carefully details how this was accomplished and lists some of the obstacles encountered with the difficult to express proteins.  As a proof of concept, a protein array was constructed using a dot blotting device and nitrocellulose sheets, which served as the solid phase platform.  The arrays were spotted and exposed to sera from Johne’s disease cattle as well as mice and rabbits that had been immunized with live M. paratuberculosis.  Antigens were readily identified in all hosts examined and these studies provided motivation to continue with the time investment required to develop more comprehensive protein arrays.
                                Technical Abstract: As an initial step toward systematically characterizing all antigenic proteins produced by a significant veterinary pathogen, 43 recombinant Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) expression clones were constructed, cataloged and stored.  Nitrocellulose filters were spotted with purified proteins from each clone along with a whole cell lysate of M. paratuberculosis.  Spots on the resulting dot array consisted of hypothetical proteins (13), metabolic proteins (3), cell envelope proteins (7), known antigens (4) and unique proteins with no similarity in public sequence databases (16).  Dot blot arrays were used to profile antibody responses in a rabbit and mouse exposed to M. paratuberculosis as well as in cattle showing clinical signs of Johne’s disease.  The M. paratuberculosis heat shock protein DnaK, encoded by open reading frame MAP3840 and a membrane protein MAP2121c, were identified as the most strongly immunoreactive in both the mouse and rabbit hosts, respectively.  Whereas MAP3155c, which encodes a hypothetical protein, was most strongly immunoreactive in Johne’s disease cattle.  This study has enabled direct comparisons of antibody reactivity for an entire panel of over 40 proteins and laid the foundation for future high throughput production and arraying of M. paratuberculosis surface proteins for immune profiling experiments in cattle.


Return to top


Connect with ARS Sign up


ARS Home
                            USDA.gov
                            Plain Writing
                            Policies & Links
                            Civil Rights Statements
                            FOIA
                            Accessibility Statement
                            Privacy Policy
                            Non-Discrimination Statement
                            Quality of Information
                            USA.gov
                            WhiteHouse.gov