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
 Molecular Plant Pathology Laboratory: Beltsville, MD
 close



    Research

                    Research Projects
                                    Publications
                                    Collaborations
                                    News
                People

                    Staff Listing
                                    Careers


        


                    Databases
                        Alfalfa Transcription Factor Database
                                    iPhyClassifier - Phytoplasma Sequence ClassificationSearch small
                    Search
            
            ARS Home
                                About ARS
                                Contact Us




ARS Home Northeast Area Beltsville, Maryland (BARC) Beltsville Agricultural Research Center Molecular Plant Pathology Laboratory Docs







Maize Rayado Fino Virus-VLPs and peptide crosslinkingheadline bar **Maize Rayado Virus-VLPs and peptide crosslinking on the viral surface** */ARSUserFiles/80420560/image_6_smaller.jpg*   *Figure legend: Two step protein cross-linking procedures between Cys 2-VLPs and F and HN peptides. (A) Cross-linking of NHS-PEG4-Maleimide with amine groups (N-terminal) of F and HN peptides. (B) Cross-linking of peptide-NHS-PEG4-Maleimide with thiol group Cys 2-VLPs.* ** *Reference* *Natilla, A**,Hammond RW,**Maize rayado fino virus-like particles expressed in tobacco plants: a new platform for cysteine selective bioconjugation peptide display**-**J Virol Methods.**2011 Dec;178(1-2):209-15. doi: 10.1016/* **  *Maize rayado fino virus*(MRFV) (genus*Marafivirus*, family*Tymoviridae)*iscomposed of isometric particles of 30 nm in diameter. Each particle has 180 copies of the capsid protein (CP) forming a T=3 structure, stabilized by protein-protein interactions. MRFV CPs self-assemble in*Escherichia coli*and in*Nicotiana benthamiana*into virus-like particles (VLPs) identical in size and shape to MRFV native capsids, a very desirable attribute in nanotechnology (Hammond and Hammond, 2010). Our laboratory investigatedMRFV- VLPs produced in tobacco plants for their ability to serve as platform to which different peptides could be covalently displayed.  To provide an anchor for chemical modifications, we created Cys-MRFV-VLPs mutants by substituting several of the amino acids present on the shell of the wild-type MRFV-VLPs with cysteine residues. The mutant designated Cys 2-VLPs exhibited, under native conditions, cysteine thiol reactivity in bioconjugation reactions with a fluorescent dye. Successively, Cys 2-VLPs was cross-linked by NHS-PEG4-Maleimide to 17 (F) and 8 (HN) amino acid long peptides, corresponding to neutralizing epitopes of*Newcastle disease virus*(NDV). The resulting Cys 2-VLPs-F and Cys 2-VLPs-HN were recognized in Western blots by antibodies to MRFV as well as to F and HN.  The results demonstrated that plant-produced MRFV-VLPs have the ability to function as a novel platform for the multivalent display of surface ligands.


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