Expression and Purification of ATM Protein using Vaccinia Virus
BACKGROUND: Ataxia telangiectasia (AT) is a genetic recessive disorder involving a large range of symptoms including telangiectasia (dilation of blood vessels) on the eyes, face, and shoulders, ataxia (loss of balance), cerebellar degeneration, radiosensitivity, cancer predisposition, immunodeficiency and premature aging. At a cellular level, AT cells display cell cycle checkpoint defects, chromosomal instability and sensitivity to ionizing radiation. The protein mutated in AT, ATM (Ataxia Telangiectasia-Mutated), is a large protein of 370 kDa and is very difficult to purify. All previous efforts (using bacterial, yeast, and baculovirus systems) have failed to achieve a meaningful level of protein expression. Purified protein is important not only to further research regarding biochemical and functional characterization of this protein, but is also crucial for development of diagnostic assays.
DESCRIPTION: Researchers at the University of California have developed a method for recombinantly producing a high yield of functional ATM protein. Specifically, they have generated a recombinant vaccinia virus containing ATM that can be used to infect mammalian cells. Once infected, the cells produce large amounts of ATM protein, which is then purified from the cells.
* The vaccinia virus expression system can be used to purify ATM protein at substantially higher levels than possible using previously reported methods.
* The new method is further scalable, allowing production of large amounts of ATM protein.
* The ATM protein generated using this method is fully functional.
* Purified ATM protein can be used for research purposes requiring biologically active protein, such as for biochemical and functional characterization of the protein.
* Purified ATM protein can be used for the development of diagnostic assays.
US 6,994,975 [MORE INFO
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