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XMRV Protease Plasmid

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Proteins and Proteomics

ATP

Page last updated February 11, 2011

XMRV Protease Plasmid

The clones for XMRV protease, as well as clones for

all of the other protein products encoded by XMRV,

were produced by Protein Expression Laboratory

(http://bit.ly/eqz7ZT) to support an NCI effort to

develop standard reagents to study this newly

discovered human virus implicated in prostate cancer

and chronic fatigue syndrome.

In the case of XMRV protease, there were multiple

clones produced and this was a critical asset to this

recent paper.

*It was through using an MBP-protease fusion

protein that we had the first indication that the lab

was working with an enzymatically active protein,*

says Dr. Wlodawer.

The structural findings suggest that the XMRV

protease may be a member of an evolutionary

distinct branch of aspartic proteases.

The clone development project in the PEL was

headed by Dr. Dom Esposito with technical support

from Wall and Mehalko.

*The project to clone all the XMRV-encoded protein

products was an NCI priority in the laboratory*

says Dr. Esposito.

The cloning work took less than 4 weeks to carry out,

and generated more than 40 different expression

clones for use in producing protein from multiple host

organisms ranging from bacteria to mammalian

cells.

The clones produced by the PEL have been used in a

variety of ways in addition the protein structure work

of Dr. Wlodawer's laboratory.

Proteins were purified at the PEL for use in

development of serological assays for XMRV

infection, and were also used to generate

monoclonal antibodies.

In addition, clones for various proteins were

distributed to researchers across the country for

biochemical studies, and are now freely available

from the AIDS Reference Reagent Repository

(http://bit.ly/dLAKa2).

These clones were generated using the Gateway

recombinational cloning system, which allows very

simple transfer of the protein-coding regions to

different vectors for many different purposes.

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ATP - Advanced Technology Program

The Advanced Technology Program (ATP) provides

cutting-edge core services through highly developed

expertise and technologies to help the National

Cancer Institute (NCI) and other NIH institutes and

government agencies meet the challenges in

biomedical research.

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