= History and Background =
In 1978 scientists were just beginning to understand the potential of what Boyer and Cohen had done by cutting DNA into specific fragments using restriction enzymes, rejoining them and transferring them into bacterial host cells [ [http://web.mit.edu/invent/iow/boyercohen.html Lemelson-MIT Program (1997) Stanley Cohen and Herbert Boyer. "Inventor of the Week".] (accessed July 7, 2008) ] . Promega Corporation founder and CEO, Bill Linton, realized that if researchers had these enzymatic tools readily available to them, they would have more freedom to focus on their research question: “What does that piece of DNA do in the cell?”
The industry of biotechnology got its start with Boyer and Cohen’s patents on recombinant DNA technology. Promega Corporation got its start in Bill Linton’s laboratory in 1978 with the isolation of a few restriction enzymes [ [http://www.promega.com/enotes/features/fe0016_tabs.htm Promega Corporation (2004) 25 Years. eNotes] (accessed July 9, 2008)] .
Promega is a well respected, forward-looking member of the biotechnology industry and now offers more than 2,000 life science products to research scientists in the fields of genetic identity/forensics, proteomics, molecular diagnostics, cellular analysis, drug discovery, and of course, genomics [http://www.madison.com/archives/read.php?ref=/wsj/2008/05/25/0805250223.php Newman, J. (2008) Madison biogiant: Promega helped put Madison on the world's biotechnology stage. "Wisconsin State Journal". May 24. ] (accessed July 7, 2008)] .
The privately held company has been profitable since 1984 and has its headquarters in Madison, WI, USA. Its global reach includes branch offices in 13 countries and manufacturing facilities in San Luis Obispo, California, USA; Shanghai, China; and Seoul, South Korea. From 2006 to 2007, company revenues grew by more than 10% to 220 million dollars (US) with approximately 12% reinvested in research and development [http://wistechnology.com/articles/3892/ Vanden Plas, J. (2008) Promega Corporation: Wisconsin's quiet innovator. "WTN News" May 2.] (accessed July 1, 2008)] . Promega Corporation also established the first biotechnology joint venture in China (Sino-American Biotechnology Co. in 1985) [ [http://www.madison.com/archives/read.php?ref=/wsj/2008/05/25/0805250226.php Newman, J. (2008) Promega, not quite your average U.S. work space. "Wisconsin State Journal" May 24.] (accessed July 1, 2008) ] .
As of 2008, 145 US patents were assigned to Promega Corporation, and the company holds a number of foreign patents as well [ [http://patft.uspto.gov/ United States Patent and Trademark Office Web site search for Patents Assigned to Promega Corporation] (July 1, 2008) ] . In addition to developing its own intellectual property, Promega works with academic institutions and other entities to license and develop promising technologies. As a member of the Wisconsin Alumni Research Foundation Research Tool Subscription Program, Promega has the opportunity to take a first look at new technologies from the university [ [http://wistechnology.com/articles/2299/ 2005. Promega to get early access to WARF technologies under new agreement. "WTN News". Sept. 28.] (accessed July 7, 2008).] .
Promega strives to be an industry leader not only in product innovation, but also in methods of delivery to laboratory scientists, such as the Promega"Express"™ on-site stocking system, which uses radio frequency identification (RFID) linked to the internet to track and manage remote inventory. This technology ultimately resulted in the spin-off company [http://www.tersosolutions.com Terso Solutions] that specializes in the design and manufacture of small RFID storage units [ [http://www.madison.com/archives/read.php?ref=/tct/2005/11/15/0511150112.php (2005) Report: Promega spins off RFID unit. "The Capital Times". November 15. Business, 8B.] (accessed July 7, 2008).] .
= Product Areas and Technologies =
In the field of genomics, Promega DNA and RNA isolation systems, cloning vectors, and site-directed mutagenesis systems are popular products. A search of the HighWire® database (July 2007–2008) for the Promega T-vector cloning product “pGEM®-T” produced over 1200 citations [pGEM®-T is a vector that allows direct cloning of products generated by polymerase chain reaction. Such products often have adenine overhangs on their ends, and can be directly cloned into a vector with T (thymine) overhangs. The original pGEM®-T vector was introduced by Promega in 1992.] . Promega also has a large portfolio of amplification products, including the GoTaq® family of polymerases and buffers and the Plexor® quantitative PCR system. In addition, Promega has recently introduced a 10-minute plasmid miniprep system that yields transfection-quality DNA for cell biology applications [ [http://www.promega.com/cnotes/cn021/cn021_03.pdf Smith, D. and Vincent, E. (2008) Transfection-quality plasmid DNA in as little as ten minutes using the PureYield™ Plasmid Miniprep System. "Cell Notes". 21, 3–5.] ] .
Genetic Identity and Forensics
Promega is one of two main companies worldwide that provide systems to scientists for genetic identification based on DNA analysis using short tandem repeats (STRs), [http://dna.gov/ Amplification on-line training course at DNA.gov] (Note: The amplification course requires registration)] .
DNA-based human identification or DNA finger-printing was originally developed by Sir Alec Jefferys, who identified repetitive regions of human genomic DNA. These regions vary in length based on the number of repeats (variable number of tandem repeats, VNTR) [http://www.promega.com/profiles/1002/ProfilesinDNA_1002_03.pdf Donelly, S. (2007) An interview with Sir Alec Jefferys. "Profiles in DNA" September. 3–5.] (accessed July 14, 2008).] [http://www.dna.gov/basics/analysishistory/ History of DNA Analysis] (accessed July 14, 2008)] . Individuals possess different numbers of repeats in these sequences, which can be detected on gels and used as an unique identifier for an individual. VNTRs were used in an immigration case in 1985; since that time improvements in DNA isolation and detection have made DNA-based identification a standard in forensic and medical laboratories.
Current DNA-based identification methods are based on PCR amplification of STR loci. Promega was the first company to provide kits for STR analysis of single loci. Along with Applied Biosystems, Promega participated with the FBI and other crime labs in validating STR loci that would eventually be selected as the core loci for the COmbined DNA Index System (CODIS), used for forensic DNA testing in North America.
The Promega PowerPlex® STR systems were the first commercially available systems for STR analysis that contained all of the CODIS loci. These systems have been used to identify the remains of victims found in mass graves from wars in Croatia, Bosnia and Herzegovina in the early 1990s [ [http://www.cmj.hr/2001/42/3/11387635.pdf Alonso, A. "et al". (2001) DNA Typing from skeletal remains: Evaluation of multiplex STR systems on DNA isolated from bone and teeth simples. "Croatian Medical Journal '42", 260–6.] (accessed July 7, 2008)] . Promega also sent scientists to New York immediately after the September 11, 2001, attack on the World Trade Center to assist in identifying victims .
The development of eukaryotic cell-free translation systems based on rabbit reticulocyte lysate (RRL) was a major advance for studying protein functionArduengo, M., Schenborn, E. and Hurst, R. (2007) The Role of Cell -Free Rabbit Reticulocyte Expression Systems in Functional Proteomics. In: "Cell-Free Protein Expression". Kudlicki, W. "et al". (ed.) Landis Bioscience (Austin, TX). ISBN 978-1-58706-123-3] . Promega Corporation was an early supplier in the cell-free protein synthesis field and is continuing to develop its portfolio in this area [http://dx.doi.org/10.1007/s10969-007-9035-2 Zhao, K.Q. "et al". (2007) Functional protein expression from a DNA-based wheat germ cell-free system. "J. Struct. Funct. Genomics". 8, 199–208.] ] [ [http://www.promega.com/pnotes/ Leippe, D. (2008) Cell-Free protein expression with the TnT® T7 Insect Cell Extract Protein Expression System. "Promega Notes" 100, 11–12.] ] . Since the introduction of the original RRL systems, coupled transcription and translation systems, which use DNA rather than the more labile RNA as the template, have become available. Additionally, researchers now have the option of using nonradioactive methods to detect and capture proteins, and they can synthesize proteins on a large or small scale, depending on their experimental needs [ [http://www.promega.com/cnotes/cn021_6.pdf Kobs, G. (2008) Selecting the cell-free protein expression system that meets your experimental goals. "Cell Notes 21", 6–9.] (accessed July 17, 2008)] .
The sequencing of the human genome in 2003 ushered in an era of high-throughput proteomics. With the gene sequences readily available, researchers’ questions turned to the function of protein-coding sequences within the genome and the interactions between protein networks driving cellular processes. For example, Promega TnT® systems have been used in a high-throughput protein truncation test-ELISA screen for mutations of the ataxia-telangiectasia mutated (ATM) gene that disrupt full-length protein synthesis [ [http://dx.doi.org/10.1016/j.mrfmmm.2008.01.002 Du, L. "et al". (2008) Rapid screen for truncating ATM mutations by PTT-ELISA. "Mutat. Res". 640, 139–44.] ] . Studying networks of proteins requires expression of large numbers of proteins from expansive cDNA libraries. The Promega Flexi® Vector Cloning System was used in constructing both the KDRI HUGE collection of full-length cDNA clones and the commercially available Origene FlexClone collection [ [http://dx.doi.org/10.1093/hmg/ddl048 Temple, G. (2006) From genome to proteome: Developing expression clone resources for the human genome. "Hum. Mol. Genet." 15, R31–R43] ] .
The Promega HaloTag® Technology Platform is a technical advance for protein study. The HaloTag® protein is a modified halogenase that forms a covalent bond with synthetic ligands. [http://dx.doi.org/10.1021/cb800025k Los, G. "et al." (2008) HaloTag: A novel protein labeling technology for cell imaging and protein analysis. "ACS Chem. Biol. 3, 373–82.] ] . A researcher “tags” a protein of interest by creating a fusion with the HaloTag® protein. Then one of several ligands can be used to visualize, capture, and track proteins. HaloTag® technology is being incorporated into commercially available open reading frame libraries to create ready-to-use expression clones [ [http://www.drugdiscoverynews.com/index.php?newsarticle=2237 Anderson, C. (2008) Promega and GeneCopoeia play tag. Drug Discovery News. June.] (accessed July 17, 2008)] .
Cellular Analysis and Drug Discovery
Promega offers a range of products for cellular analysis including reporter assays, apoptosis detection systems, cell-signaling assays, and cell-based assays. Many of these systems and reagents are built upon Promega bioluminescence technology.
Cell-based assays that use bioluminescence eliminate many problems associated with overlap between test compound excitation/emission wavelengths and fluorescent reporters, which can confound assay results [http://dx.doi.org/10.1089/adt.2006.053 Fan, F. and Wood, K.V. (2007) Bioluminescent assays for high-throughput screening. "ASSAY and Drug Development Technologies". 5, 127–36.] ] . Luciferase is not endogenous to mammalian cells, and therefore inherent background is reduced, dramatically increasing assay sensitivity. This sensitivity allows researchers to conserve precious cell samples and detect signals from a subset of a mixed cell population [ [http://www.promega.com/cnotes/cn020/cn020_06.pdf Capowksi, E.E. and Svendsen, C.N. (2008) Monitoring cell viability during gene directed enzyme prodrug therapy with the CellTiter-Glo® Assay and human neural progenitor cells. "Cell Notes 20", 6–8.] ] .
One of the most recognized bioluminescence-based assays is the Dual-Luciferase® Reporter Assay, which allows researchers to differentiate changes in signal that are biologically relevant from nonspecific changes through normalization to a control. Such dual-luciferase reporter assays are used to monitor activity of G-protein coupled receptor pathways, investigate modulation of gene activity through antioxidant response elements [ [http://dx.doi.org/10.1124/mol.107.042499 Wruck, C.J. "et al". (2008) Kavalactones protect neural cells against amyloid-β peptide-induced neurotoxicity via ERK 1/2-dependent Nrf-2 activation. "Mol. Pharmacol.". 73, 1785–95.] ] , and identify molecules that enhance STAT-1-dependent gene expression in a high-throughput screen [ [http://dx.doi.org/10.1158/0008-5472.CAN-06-2439 Lynch, R.A. "et al". (2007) A small-molecule enhancer of signal transducer and activator of transcription 1 transcriptional activity accentuates the antiproliferative effects of IFN-γ in human cancer cells. "Cancer Research 67", 1254-61.] ] .
By modifying the bioluminescence approach so that ATP or luciferin rather than luciferase is the limiting reagent, Promega has developed assays that allow researchers to measure parameters other than gene activity. For instance, assays for kinase inhibitors are of particular interest to the drug discovery community, and luminescent assays such as the Kinase-Glo® Assays provide a universal screening method for identifying inhibitors using virtually any kinase and substrate combination.
Additionally, since ATP is a marker of cell viability, ATP assays are used to detect microbes and quantify mammalian cells. Bioluminescent ATP-based cell viability assays, such as the CellTiter-Glo® Assay, are more sensitive than methods that assess redox potential in cells, and additionally the CellTiter-Glo® Assay requires only minutes (compared to hours) to perform. The widespread use of bioluminescence-based technologies for screening is demonstrated by a search of the PubChem BioAssay database, which has 131 entries that describe Promega technologies or assays used by MLSCN screening centers [The [http://www.ncbi.nlm.nih.gov/sites/entrez?holding=wipmglib PubChem BioAssay Database] was searched for the keyword “promega” and returned 131 results. (July 18, 2008)] .
Biological probes and drug candidates identified during screening must be characterized further for adsorption, deposition, metabolism excretion and toxicology (ADME/Tox). Traditionally, ADME/Tox assays are performed in animal models, but animal testing is expensive, raises moral questions, and may not be truly predictive, so drug developers are turning to cell-based and in vitro assays [ [http://www.dddmag.com/adme-tox-gets-faster-cheaper.aspx Dove, A. (2007) ADME-Tox gets faster, cheaper and safer. "Drug Discovery and Development." September 24] . (accessed July 18, 2008)] . Bioluminescence-based screening technologies may help drug developers eliminate toxic compounds earlier in the development process, reducing failed clinical trials or after-market recalls [http://www.genengnews.com/articles/chitem.aspx?aid=2509 Lipp, E. (2008) Tackling drug-interaction issues early on. "Genetics Engineering News" 28, 12.] (accessed July 18, 2008)] [Cali, J.J. (2008) Bioluminescent assays for ADMET. "Expert Opin. Drug Metab. Toxicol". 4, 103–20.] [ [http://dx.doi.org/10.1517/17425255.2.4.629 Cali, J.J. (2006) Luminogenic cytochrome P450 assays. "Expert Opin. Drug Metab. Toxicol." 2, 629–45.] ] .
Cytochrome P450 (CYP450) enzymes are involved in metabolism and clearance of many drugs and therefore are involved in toxicity and drug-drug interactions (DDI). In the P450-Glo™ Assays a luminogenic substrate is metabolized by a CYP450 to a luciferin derivative, which then reacts with luciferase to produce light.
By combining luciferase reporter gene assays and luminogenic P450 assays, researchers can predict induction of cytochrome P450 activity at the level of gene transcription and enzyme activity. This approach was useful for understanding the harmful interation between grapefruit juice and statins. A component of grapefruit juice, bergamottin, upregulates the gene encoding the cytochrome P450 that metabolizes the cholesterol-lowering statins, but at the same time, bergamottin inhibits the activity of the actual enzyme, slowing statin clearance.
Promega bioluminescence assays, DNA and RNA purification chemistries, and HaloTag® technologies integrate with the high-throughput automated systems found in many laboratories. Some of this integration has occurred through collaboration with instrument manufacturers, with Promega providing the chemistry expertise.
Promega also sells the Maxwell® 16 System, which is a bench-top automated purification system for lower throughput research and diagnostic laboratories. The Maxwell® 16 System purifies DNA, RNA, and recombinant protein and processes up to 16 samples in 30–45 minutes, depending on sample type. This system has been used to purify proteins for structural studies [Frederick, R. (2007) [http://www.uwstructuralgenomics.org/presentations/Bottlenecks_07_frederick_slides.pps Three-part, small-scale screening platform for the masses.] Center for Eukaryotic Structural Genomics. Department of Biochemistry. University of Wisconsin Madison. PowerPoint® Presentation (accessed July 18, 2008)] and to purify DNA from a variety of sample types and organisms [ [http://portal.knowledgebase.net/display/1/index.asp?c=10537&cpc=WnyQDcP53qMu523S1NA631wwrR4UxgK6fv2ylkXWS&cid=11781&cat=&catURL=&r=0.765774 Maxwell® Applications Database] (accessed July 18, 2008)] .
Promega GloMax® Luminometers are supplied with preinstalled protocols that allow researchers to perform multiplex bioluminescent assays. The luminometers with injection systems are available for use with dual-reporter assays like the Dual-Luciferase® systems.
The Promega Maxwell® 16 System is compliant as a General Purpose Medical Device in the United States and has received the CE Mark for diagnostic use throughout the European Union [(2008) [http://www.genomeweb.com/issues/news/147222-1.html Promega gets CE Mark for DNA purification system. "GenomeWeb Daily" May 29.] (accessed July 18, 2008)] . This bench-top, personalized approach to automation allows clinical labs to benefit from more reproducible results and improved work flow with easy installation and minimal training [Berman, R. "et al." (2007) Automated sample prep to improve lab workflow. "Automation for diagnostics: IVD Technology Supplement" Sept. 11.] [Thomson, A. (2007) Insights into lab automation’s future. "IVD Technology" January, 66.] .
The Y-Chromosome Deletion Detection System from Promega also carries the CE Mark for use as an in vitro diagnostic device in the European Union.
= Manufacturing =
Promega was first certified to international standards for quality management systems in 1998 and is currently certified to the ISO 13485 standard, required for the development, manufacture, testing, and delivery of medical devices around the world. The ISO series of quality management system standards are developed and maintained by the International Organization for Standardization. An organization achieving ISO certification has demonstrated to a third-party body of experts that the organization meets all requirements of the standard.
= Corporate Citizenship and Environmental Record =
According to Jim Leonhart, Executive Director of the Wisconsin Biotechnology and Medical Device Association, Promega Corporation is not only a leader in the biotechnology industry, but it is also a “good corporate citizen” (3). When Promega broke ground for the BioPharmaceutical Technology Center (BTC) in 1994, 20,000 square feet of that building was devoted to community use, including a 300-seat auditorium, conference rooms, and teaching labs that are part of the [http://www.btci.org/ BioPharmaceutical Technology Center Institute] (BTCI), a nonprofit organization of which Promega is the founder and main corporate sponsor [ [http://www.madisonmagazine.com/article.php?section_id=918&xstate=view_story&story_id=125828 Howell, B. (2003) The science of cool. "Madison Magazine".] (accessed July 2, 2008)] [http://www.btci.org/ BioPharmaceutical Technology Center Institute] (accessed July 7, 2008)] . Activities hosted by BTCI include the Annual International Bioethics Forum, the Stem-Cell Symposium, and the laboratory course for the Youth Apprenticeship Program in Biotechnology.
In addition to the outreach provided through the BTCI, Promega runs a [http://www.promega.com/us/trainingsupport/ Training Support Program] that provides Promega products at significant discounts to educators in the US and provides [http://www.promega.com/education/ free educational resources] for undergraduate instructors. The [http://www.fitchburgcenter.com/artintheatrium.htm BTC Atrium Gallery] also hosts quarterly art shows that feature local artists.
Promega Corporation has a reputation for environmental responsibility. In 1994, when the BTC was built, Promega used native prairie landscaping to reduce use of herbicides and fertilizers and sustain wildlife habitat [ [http://www.prairienursery.com/services/promega_corp.htm Prairie Nursery Case Study] .(accessed July 7, 2008)] . The Agora, which opened in November 2006, is the focal point of Fitchburg Center and the headquarters for Promega Corporation. This building retains Promega’s commitment to environmentally friendly design. The building has a storm water collection system that allows runoff to drain into the adjacent prairie for natural filtering and underground parking to minimize the amount of asphalt and concrete associated with development of the area [Ivey, M. (2006) [http://www.madison.com/archives/read.php?ref=/tct/2006/09/14/0609140311.php Written in Stone: Fitchburg town center embraces environment. "The Capital Times." Sept. 14.] (accessed July 7, 2008)] .
Additionally, Promega practices environmentally responsible shipping and packaging practices, reducing disposable packaging materials whenever possible and using recycled materials for packaging and print materials. For large shipments to Europe, Promega uses Enviro Containers.
= Footnotes =
= External Links = [http://www.promega.com Promega Corporation]
[http://www.btci.org BioPharmaceutical Technology Center Institute]
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