G protein-coupled receptors (GPCRs) are a large family of receptors that traverse the cell membrane seven times. Functionally, GPCRs are extremely diverse, yet they contain highly conserved structural regions. GPCRs respond to a variety of signals, from small molecules to peptides and large proteins. Many GPCRs are involved in disease pathways and, not surprisingly, they present attractive targets for both small-molecule and biologic drugs.
In response to a signal, GPCRs undergo a conformational change, triggering an interaction with a G protein—a specialized protein that binds GDP in its inactive state or GTP when activated. Typically, the GPCR exchanges the G protein-bound GDP molecule for a GTP molecule, causing the activated G protein to dissociate into two subunits that remain anchored to the cell membrane. These subunits relay the signal to various other proteins that interact with or produce second-messenger molecules. Activation of a single G protein can result, ultimately, in the generation of thousands of second messengers.
Today’s blog brought to you by Julia Nepper, a Promega science writer guest blogging for the BioPharmaceutical Technology Center Institute (BTC Institute)!
“We all benefit from STEM role models. When students from underrepresented populations meet and learn about STEM professionals of color, they can see themselves as the scientists and engineers of the future. Fun, engaging science programming for children is also essential to light the spark for the next generation. A Celebration of Life, the partnership between the BTC Institute and the African American Ethnic Academy, two community nonprofits, has combined these 2 objectives for over twenty years.” according to Barbara Bielec, K-12 Program Director.
This year, the theme of the program is Sunsational!, with a number of activities related to the sun, solar energy, and STEM careers. As part of the program, students heard talks from several STEM professionals of color about their work. Mehrdad Arjmand, co-founder of solar energy company NovoMoto, was one of those speakers.
Dr. Arjmand was born and raised in Iran. His path to becoming a mechanical engineer began as a child, with him “destroying a lot of equipment” in his house. After completing his undergraduate education, he came to the States to pursue a PhD at the University of Wisconsin-Madison, where he met Aaron Olson, a student who was born in the Democratic Republic of Congo. These two discovered a shared passion for starting a business and helping their communities, which led directly to the founding of NovoMoto. The name derives from Portuguese for “new” (novo) and Lingala—a language spoken in Congo—for “fire” (moto). Continue reading “Empowering Communities with the Light of the Sun”
This past May (2019) the symposium “Psychedelic Therapy in Society: Exploring the Mechanisms of Action and Delivery of Care” was hosted by the International Forum on Consciousness at the BioPharmaceutical Technology Center in Madison, WI.
Having the good fortune to work across the street at Promega, I was able to attend this two-day conference and learn from leading researchers in psychedelics and about their use in therapy.
My interest in psychedelics is relatively new. I didn’t experiment with these substances during high school or college years. But in recent years, I’ve seen a close relative struggle with profound anxiety related to terminal disease, and another with substance abuse and depression. The lessons learned from each experience is that the battery of medicines used to treat such illness can result in additional problems for which there are currently not good medication options. And in some cases, traditional medications can cause further health problems. Continue reading “Psychedelics as Therapeutic Agents: Current Research, Potential Benefits”
Restriction enzymes recognize short DNA sequences and cleave double-stranded DNA at specific sites within or adjacent to these sequences. These enzymes are the workhorse in many molecular biology applications such as cloning, RFLP, methylation-specific restriction enzyme analysis of DNA, etc. In order to streamline and shorten these workflows, restrictions enzymes with enhanced capabilities are desirable.
A subset of Promega restriction enzymes offer capabilities that include rapid digestion of DNA in 15 minutes or less, ability to completely digest DNA directly in the GoTaq® Green Master Mix, and Blue/White Cloning Qualification which allows for rapid, reliable detection of transformants.
To learn more about restriction enzymes and applications, check out Restriction Enzyme Resource on the web. The resource provides everything from information on restriction enzyme biology to practical information on how to use restriction enzymes. This resource also contains useful online tools to help you use enzymes more effectively. It helps you choose the best reaction buffer for double digests, find the commercially available enzyme that cuts your sequence of interest, find compatible ends, and search for specific information on cut site, overhang isoschizomers and neoschizomers by enzyme name.
For added convenience, you can download the mobile app and use the Restriction Enzyme Tool to plan your next digest.
For additional information regarding Restriction Enzyme Digest, reference the supplementary video below.
Some thermostable DNA polymerases, including Taq, add a single nucleotide base extension to the 3′ end of amplified DNA fragments. These polymerases usually add an adenine, leaving an “A” overhang. There are several approaches to overcome the cloning difficulties presented by the presence of A overhangs on PCR products. One method involves treating the product with Klenow to create a blunt-ended fragment for subcloning. Another choice is to add restriction sites to the ends of your PCR fragments. You can do this by incorporating the desired restriction sites into the PCR primers. After amplification, the PCR product is digested and subcloned into the cloning vector. Take care when using this method, as not all restriction enzymes efficiently cleave at the ends of DNA fragments, and you may not be able to use every restriction enzyme you desire. There is some useful information about cutting with restriction sites close to the end of linear fragments in the Restriction Enzyme Resource Guide. Also, some restriction enzymes require extra bases outside the recognition site, adding further expense to the PCR primers as well as risk of priming to unrelated sequences in the genome. Continue reading “A Quick Method for A Tailing PCR Products”
Scientific investigation is an iterative process, for which reproducibility is key. Reproducibility, in turn, requires accuracy and precision—particularly in measurement. The unsung superheroes of accuracy and precision in the research lab are the members of your local Metrology Department. According to Promega Senior Metrologist, Keela Sniadach, it’s good when the metrology department remains unsung and behind the scenes because that means everything is working properly.
Holy Pipettes, Scientists! We have a metrology department?! Wait…what’s metrology again?
Metrology (the scientific study of measurement) got its start in France, when it was proposed that an international length standard be based on a natural source. It was from this start that the International System of Units (SI), the modern metric system of measurement, was born.
Metrology even has its own day: May 20, which is the anniversary of the day that the International Bureau of Weights and Measures (BIPM) was created by the Meter Convention in Paris in 1875. The job of BIPM is to ensure worldwide standards of measurement.
Concepcion Sanchez-Cid didn’t know she wanted to be a scientist when she was older. She grew up with a love of music and played the violin, but her curiosity and eagerness to learn drove her down the path for a career in biomedical research.
Hear more of Concepcion’s story:
As a Master’s student at the University of Granada, Concepcion studied biotechnology and landed an internship at the Promega Europe Training and Application Lab (PETAL) in France. She worked with the Applications Team to develop protocols for DNA and RNA extraction from soil. When she decided to pursue a PhD, she received a sponsorship from Promega and enrolled as a student at the University of Lyon while also remaining an employee at PETAL.
Concepcion says that the balance between both worlds—academia and industry—provide her with technical skills and a unique support network that has helped shape her PhD thesis work. “Working at a university and a company at the same time…you get very different feedback from people that are very specialized, and they really know what they’re doing, so at the end you integrate everything,” she says. “It’s one of the things I appreciate most about my PhD.” Continue reading “Curiosity and Collaboration: A PhD Journey”
On May 13, 2019, twenty-five meters below the streets of Stockholm in a retired nuclear reactor, Nerea Capon and her iGEM team unveiled an artistic fusion of creativity and synthetic biology. The Synthetic Biology Art Exhibition featured works by other iGEM teams and local artists, all presenting their unique reflections on the concepts of synthetic biology. The collection included synthetic skin grown by bacteria, performance art, and even a musical snail that spent the week crawling around a table full of plants.
Innate immunity, the first line of immune defense, uses a system of host pattern recognition receptors (PRRs) to recognize signals of “danger” including invariant pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs). These signals in turn recruit and assemble protein complexes called inflammasomes, resulting in the activation of caspase-1, the processing and release of the pro-inflammatory cytokines IL-1ß and IL-18, and the induction of programmed, lytic cell death known as pyroptosis.
Innate immunity and the activity of the inflammasome are critical for successful immunity against a myriad of environmental pathogens. However dysregulation of inflammasome activity is associated with many inflammatory diseases including type 2 diabetes, obesity-induced asthma, and insulin resistance. Recently, aberrant NLRP3 inflammasome activity also has been associated with age-related macular degeneration and Alzheimer disease. Understanding the players and regulators involved in inflammasome activity and regulation may provide additional therapeutic targets for these diseases.
For over a decade, obesity has been called an “epidemic”, both in the popular and scientific literature. Traditionally, the term “epidemic” is associated with a highly contagious disease that carries with it a significant risk of mortality. A comprehensive review of observational studies (1) suggested that obesity did not fit this definition, despite the use of the term in a widely disseminated report by the World Health Organization in 2002.
Regardless of the etymological fine points, the worldwide prevalence of obesity and its associated health risks are clear. These risks include type 2 diabetes, hypertension, several cancers, gall bladder disease, coronary artery disease and stroke (2). Yet, the debate over obesity and options for reducing its risks has become increasingly polarized. As a result, some health researchers are advocating a “health at every size” (HAES) approach to address the social, cultural and lifestyle implications of obesity (2).