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Purify First: Rapid Expression and Purification of Proteins from XMRV.

Discussion in 'XMRV Research and Replication Studies' started by CBS, Dec 15, 2010.

  1. CBS

    CBS Senior Member

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    Scientists in the Protein Expression Laboratory at NCI-Frederick (same NCI location as Frank and Sandra Ruscetti) have apparently made progress in accurately identifying XMRV proteins. This article is hyper-technical. I'm not in a position to evaluate it critically. I'm posting this for those more well versed in this field. It seems that they have dramatically increased their rate of success in identifying XMRV by switching the order in which they attempt to purify and express the proteins in their samples. Who even knew there was a Journal of Protein Expression and Purification? Anyone care to elaborate? Let me know if you need a copy of the full manuscript.

    It is nice (and reassuring) to see that interest in XMRV appears to be spreading across the range of biological sub-specialties.

    Protein Expr Purif. 2010 Dec 9. [Epub ahead of print]
    Purify First: Rapid Expression and Purification of Proteins from XMRV.

    Gillette WK, Esposito D, Taylor TE, Hopkins RF, Bagni RK, Hartley JL.
    Protein Expression Laboratory, SAIC-Frederick, Inc., NCI-Frederick, Frederick, Maryland 21702.
    Abstract

    Purifying proteins from recombinant sources is often difficult, time-consuming, and costly. We have recently instituted a series of improvements in our protein purification pipeline that allows much more accurate choice of expression host and conditions and purification protocols. The key elements are parallel cloning, small scale parallel expression and lysate preparation, and small scale parallel protein purification. Compared to analyzing expression data only, results from multiple small scale protein purifications predict success at scale-up with greatly improved reliability. Using these new procedures we purified eight of nine proteins from xenotropic murine leukemia virus-related virus (XMRV) on the first attempt at large scale.
     
  2. alex3619

    alex3619 Senior Member

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    Logan, Queensland, Australia
    Hi CBS, this appears to be an article about an advance in protein cloning. My take, for us, without reading the paper, is that rather than try to isolate the viral proteins (there isn't much if XMRV is the virus in question) it is easier to take pieces of DNA and create a genetically modified organism that makes it in bulk. However the point of it, for them, is more about an improvement in the technique for doing this, so that they can get more easily get large quantities of purified protein. Large amounts of pure protein pave the way for experiments and treatments that require an understanding of the protein structure and function. Having not read the paper I could be wrong about much of this.

    You might consider putting this paper in the library or post details of where it can be found.

    Bye
    Alex
     
  3. Cort

    Cort Phoenix Rising Founder

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    Raleigh, NC
    It's nice to see them use XMRV first with their new technology. It sounds like a major accomplishment.
     

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