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Description
The coronavirus envelope (E) protein is a structural component of all coronavirus. Obtaining purified E protein in an efficient, clean, and reliable manner is needed for ongoing studies. Toward this goal the E protein of Mouse Hepatitis Virus (MHV) coronavirus was tagged with with either 6 or 10 histidine (his) residues which can be used for affinity chromatography purification. The his-tags were introduced by PCR into a cDNA of the MHV A59 virus strain at the carboxy end of the E gene. A reverse genetics approach was then used to assemble three full-length cDNAs of the viral genome, two modified with the hig-tags and a control wild-type (WT) without a tag . Full-length genomic RNAs were transcribed and electroporated into baby hamster kidney cells that express the MHV receptor (BHKR) and L2 rat lung cells. Virus was recovered after 72 h only from the 6X his-tagged genome and the WT control. Western blotting using antibodies against the E protein or the nucleocapsid (N) protein was performed after cells were infected with the recovered WT and 6X-tagged recombinant viruses. The E protein was not detected with the E antibody, but was detected with a histidine probe was used to detect the histidine residues. This indicates that the tagged protein is expressed and that the tag is present.
ContributorsHesser, Kathryn Sarah (Author) / Hogue, Brenda G. (Thesis director) / Hogue, Ian B. (Committee member) / Lim, Efram (Committee member) / Dean, W.P. Carey School of Business (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
ContributorsSchildkret, David (Conductor) / Chamber Singers (Performer) / ASU Library. Music Library (Publisher)
Created2018-02-10
ContributorsGlenn, Erica (Conductor) / Evans, Bartlett R. (Conductor) / Oh, Eun-Mi (Conductor) / Thompson, Jason D. (Conductor) / Schildkret, David (Conductor) / Concert Choir (Performer) / Arizona Statesmen (Performer) / Women's Chorus (Performer) / Gospel Choir (Performer) / Barrett Choir (Performer) / Chamber Singers (Performer) / Choral Union (Performer) / ASU Library. Music Library (Publisher)
Created2017-11-30
ContributorsUniversity Choirs (Performer) / ASU Library. Music Library (Publisher)
Created2000-11-16
ContributorsSchildkret, David (Conductor) / White, Jamilyn (Performer) / Krison, Danielle (Performer) / Barefield, Robert (Performer) / FitzPatrick, Carole (Performer) / Chamber Singers (Performer) / Choral Union (Performer) / Symphonic Chorale (Performer) / University Symphony Orchestra (Performer) / ASU Library. Music Library (Publisher)
Created2007-04-26
ContributorsLyne, Gregory K. (Performer) / Stutzman, Gina (Performer) / Woodgate, Lyn (Performer) / Cornner, Charles B. (Performer) / Rozukalns, Andris L. (Performer) / Women's Choir (Performer) / University Choir (Performer) / ASU Library. Music Library (Publisher)
Created1996-11-24
ContributorsCherland, Carl (Performer) / Fuller, Charles L. (Performer) / O'Brien, Robert (Performer) / Hooper, Wm. John (Performer) / Graduate Chorale (Performer) / Recital Chorale (Performer) / ASU Library. Music Library (Publisher)
Created1987-10-01
ContributorsRoueche, Michelle (Performer) / Partin, Darrell (Performer) / Wiest-Parthun, Karen (Performer) / Harvison, Emery (Performer) / Hernandez, Rene (Performer) / Foley, Laura (Performer) / Women's Choir (Performer) / ASU Library. Music Library (Publisher)
Created1994-04-21
ContributorsParthun, Karen (Conductor) / Hickman, Miriam, 1955- (Performer) / University Choir (Performer) / Vocal Jazz Ensemble (Performer) / ASU Library. Music Library (Publisher)
Created2000-11-19
ContributorsPeterson, Craig C. (Director) / Butler, Robb (Director) / Forkner, Christina (Contributor) / Leman, Christi (Performer) / Miller, Kenny (Musician) (Performer) / Howard, Johnothon (Performer) / Gibson, Ted (Performer) / Herberger Singers (Performer) / University Choir (Performer) / ASU Library. Music Library (Publisher)
Created2003-10-12