Call: 1.800.248.7609


Bookmark and Share
0 Item(s) in Cart | View Cart

Shared Results

Tagged with “Goldbio”

Posted by Chris on October 14th, 2014  ⟩  0 comments

Our modern day view of our bodies has changed quite drastically over the last century. With increased medical knowledge and the ability to detect ever-smaller and rarer life, we have continuously advanced our awareness of the world beyond that which even science fiction could have envisioned in the early 1900s. Our appreciation of life has evolved from the quaint early 20 th century belief that our bodies are pristine and pure vessels of life that are occasionally invaded by malevolent unseen forces of evil. In this modern era, we are coming to grips with the knowledge that we are fortunate symbionts, providing an ideal environment for unseen and untold communities of bacterial life who in return work tirelessly to keep us sustained, stable and productive.

As the host, it is our responsibility to maintain the proper and healthy living conditions that our bacterial tenants require. That primarily means providing timely nourishment; and our body is constructed in such a manner to remind us, like the blinking of a “low fuel” light in a car, when we need to eat or drink. As long as we are healthy, this system works and has worked spectacularly for millions of years! But when we get really sick, when our system is either severely compromised or invaded and plundered by rogue bacteria and viruses, it can be the equivalent of a Chicxulub crater-like meteor impact on the biodiversity of life within our bodies.

However, even in situations that aren’t as dire as all that, in those times when we are plagued only by the more routine dilemmas of life like the common cold, our internal stability can be compromised, albeit to a somewhat lesser extent. Throughout the millennia, our bodies have developed their systems of emergency response. One of those responses is the temporary reallocation of energy demand. When we get sick, our bodies press all available resources into fighting the infection. Consequently, that also means that our bodies aren’t worrying about finding or ingesting food. That’s a great, short-term solution…just like temporarily pulling in army reserves to bolster troops in times of need.

Just like in a prolonged war, though, that doesn’t work so well in the event of a long-term infection. Without regular and continued nourishment, the bacteria that are helping to fight the infection starve and die off, unbalancing our delicately bacterial ecosystem or allowing the existing infection to gain traction. Neither is ideal, and it makes sense that our bodies would have accounted for such an annoying dilemma with its usual flare of resourcefulness and grace. Featured in this October's issue of Nature, Joseph Pickard and Alexander Chervonsky illustrate the novel way in which mice bodies handle a similar situation.

In an ensuing infection, gut microbes need to eat even as the body is forced into starvation mode. So the body utilizes a unique system that adds a fucose sugar onto globules of fats and proteins that the bacteria can use for nourishment in times of stress. It’s not ideal and is probably similar to a military MRE (Meal Ready-to-Eat) in terms of deliciousness, but it gets the job done. In order to generate that microbial MRE, the body uses Interleukin-22 (IL22) inside of intestinal epithelial cells to express the Fut2 gene. As proof of its nourishing ability, when mice that were genetically bred to lack the Fut2 gene were stressed with a simulation of an infection, they required an extra day to return to their normal weight than the control mice.

Pickard stated in the article, the mouse fitness could either be related to decreased pathogen burden (aka resistance) or increased pathogen tolerance. To prove their hypothesis, they measured the abundance and activity of the C. rodentium bacterial infection from each test group of mice using a GoldBio L-fucose substrate called 4-Methylumbelliferyl fucopyranoside (Cat # M-580). There was no difference in the pathogen loads between the test and control groups, which showed that the Fut2 expression most likely provides beneficial tolerance as opposed to resistance to infection.

Interestingly, Pickard also noted that there is a very similar pathway that regulates antimicrobial proteins and acts as a resistance mechanism inside the body. So perhaps tolerance and resistance goes hand in hand after all. Regardless, the entire story is shaping up to be a fascinating situation and one that definitely needs further investigation. For instance, the genetic deletion of the Fut2 gene happens in 1 out of every 5 people. Fut2 has also been associated with the inflammatory bowel disorder, Crohn’s disease. Crohn’s is a terrible disease that can affect any part of the GI tract and is a life-long, debilitating condition. Crohn’s disease also happens to be genetically predisposed to run in families.

There might not be a direct link to humans yet. But it may be that without the beneficial Fut2 gene, and the timely delivery of fucose nutrients to our microbial partners, our beneficial bacteria cannot survive in the wake of intestinal infection, their loss leading to more severe physical repercussions and disabilities that otherwise might be mitigated in a healthy individual. One thing is for certain, in sickness or in health, our very lives and the quality of those lives depend on a relationship we have with organisms we cannot see, creatures we do not know, and life of which we are just now becoming aware.

I can only imagine what amazing things we will learn throughout the next century of science!

Pickard, J. M., Maurice, C. F., Kinnebrew, M. A., Abt, M. C., Schenten, D., Golovkina, T. V., ... & Chervonsky, A. V. (2014). Rapid fucosylation of intestinal epithelium sustains host-commensal symbiosis in sickness. Nature.

Category Code: 79102 88241

Posted by David on October 8th, 2014  ⟩  0 comments

In part two of our series of blogs highlighting recently published research involving the use of bialaphos and PPT, we now present a summary of exciting research utilizing Gold Biotechnology’s bialaphos and phosphinothricin (PPT) to demonstrate dramatically improved Agrobacterium-mediated ryegrass transformation.

Perennial ryegrass, or Lolium perenne, stands as the preferred cool-weather pasture and forage grass in many regions throughout the world and it serves as an important groundcover. Although perennial ryegrass is suitable for all grazing livestock, it is primarily used as the main source of nutrition for lactating dairy cows, due to the superior quality and digestibility of this species. Ryegrass is also widely utilized as a turf grass and southern state `overseed’, comprising about 40% of the nearly 50 million acres of US lawns, and it is a favorite for use in sporting fields, such as the Kansas City Royals’ Kauffman Stadium, the Seattle Mariners’ Safeco Field and golf courses worldwide.

Despite the broad cultivation of this diploid monocot, it has been quite recalcitrant to genetic modification methods, save traditional breeding, with previous transformation efficiencies reaching only about 1%.  In a recent publication, North Carolina State scientists working under a grant from Bayer Crop Science and using GoldBio’s bialaphos and PPT, reported a dramatic improvement in transformation of Lolium perenne. The researchers achieved a level of 20% transformation efficiency via cocultivation of ryegrass callus with an Agrobacterium tumefaciens strain harboring a binary vector. This plasmid contained the bar gene, which provides resistance to bialaphos and PPT, and a gene encoding green fluorescent protein (GFP).

To take this significant step forward, the researchers at North Carolina State developed a variety of modifications to traditional methods of transforming perennial ryegrass. Transformation efficiency during each experiment was measured by counting the number of GFP-expressing callus and dividing this sum by the total number of callus cocultivated with the Agrobacterium strain.

The group first demonstrated a two-fold improvement in transformation efficiency via a change in infection culture medium. The scientists employed a Murashigie and Skoog (MS)-type media, instead of the more traditional YEP infection medium, when culturing the Agrobacterium strain. The strain was then used to infect two-to-six month old ryegrass callus tissue.

Next, the team determined that a three minute 42 °C heating treatment during the initial infection with the Agrobacterium strain allowed them to achieve a four-fold increase in GFP-expressing callus selected on GoldBio’s bialaphos or PPT (Figure 2 below). Further experiments (Figure 3) showed that increasing the maltose concentration from 3% to 6% in the cocultivation medium and switching to NS6 medium (similar to MS) for that step gave an additional four-fold increase in efficiency.

Catching GoldBio Ryegrass Fig 2-3

When transgenic bialaphos-resistant GFP-expressing plants were eventually grown from callus tissue transformed using a combination of the medium modifications and heating step described above, the efficiency was shown to be more than 20%, marking a 20-fold improvement over previous efficiency in this agriculturally important plant species. This equates to an average of 20 GFP-expressing plants, like those pictured in Figure 7, per 100 callus transformed. The authors proceeded to apply this learning to transformation of rice, Oryza sativa, and observed significant improvements in efficiency with that critical crop as well.

Catching GoldBio Ryegrass Fig 7

Our third blog entry in this series will discuss the generation of bialaphos-resistant soybean, using GoldBio products, that was developed at Nanjing University. GoldBio is excited to be the choice of researchers seeking to further understanding of the biology of crop plants important to populations throughout the world.

Patel, M., Dewey, R. E., & Qu, R. (2013). Enhancing Agrobacterium tumefaciens-mediated transformation efficiency of perennial ryegrass and rice using heat and high maltose treatments during bacterial infection. Plant Cell, Tissue and Organ Culture (PCTOC), 114(1), 19-29. Plant Cell Tiss Organ Cult (2013) 114:19–29 DOI 10.1007/s11240-013-0301-7

Zhang, W. J., Dewey, R. E., Boss, W., Phillippy, B. Q., & Qu, R. (2013). Enhanced Agrobacterium-mediated transformation efficiencies in monocot cells is associated with attenuated defense responses. Plant molecular biology, 81(3), 273-286. Plant Mol Biol (2013) 81:273–286

Category Code: 88241

Posted by David on September 16th, 2014  ⟩  0 comments

Farmers are constantly in need of a more diversified herbicide treatment program. Though RoundUp has held the position of the world’s top selling herbicide for quite some time, scientists and farmers have been hard at work testing alternatives, such as bialaphos, to better handle the near-certainty and complication of herbicide tolerance. This natural herbicide, along with its metabolite phosphinothricin, marketed in its ammonium salt form as Liberty or Basta, appears to be gaining popularity. Scientists have been successfully producing bialaphos-resistant crop plants through current transfection technologies. The development of these new crops will offer farmers the flexibility of using bialaphos and phosphinothricin as effective alternate herbicides.

Briefly, as discussed in a 2012 product spotlight, bialaphos becomes phosphinothricin (PPT) in the plant cell, which inhibits glutamine biosynthesis unless transgenic enzymes encoded by the pat or bar genes are present in the plant to break it down. We will now begin a three-part series of blogs highlighting recently published research involving the use of this herbicide.

Recently, employing Gold Biotechnology’s bialaphos and PPT for selection, North Carolina State scientists, working under a grant from Bayer Crop Science, demonstrated improved Agrobacterium-mediated ryegrass crop transfection using heat and maltose treatment. Half a world away in Taiwan, GoldBio’s bialaphos was also chosen by a group of Nanjing University (PRC) scientists for use in research published this year reporting the creation of a bialaphos-resistant soybean line via Agrobacterium tumefaciens-mediated transfection. Finally, a group at Shimane University in Japan recently constructed new Gateway® binary vectors employing the bialaphos resistance (bar) gene for use in studying plant promoters. Their research focuses on powerful promoter:reporter analysis techniques to study tissue and cell-specific gene expression.

In the case of the Japanese research team’s work, R4L1 Ti plasmids for Agrobacterium-mediated transfection were developed with multiple reporter genes, such as G3GFP, G3 green fluorescent protein and GUS, B-glucoronidase in addition to selectable markers like bar, which conveys bialaphos resistance. The bar gene is under the control of the constitutive nopaline synthase (nos) promoter and terminator, while the reporter genes: G3GFP, TagRFP, GUS, etc., often linked together to be polycistronic, depend upon the host promoter adjacent to the plant chromosomal insertion site for expression. Tissue-specific promoter activity can be interrogated in robust fashion using this method. In the image below, panel F shows selection on phosphinothricin ammonium, while panel J shows imaging of GFP (green) and TagRFP (red) expression in leaf tissue. The key advantage of employing binary vectors for transfection via Agrobacterium lies in the ability to generate plant lines from plants already containing separate genetic modifications, which remain homozygous in the progeny. Previous methods involved crossing promoter:reporter plants with existing lines to stack the desired modifications. This older methodology required laborious analysis of progeny to find the desired genotypes.

Future blog entries will further discuss the generation of bialaphos-resistant ryegrass achieved at North Carolina State University and the bialaphos-resistant soybean developed at Nanjing University. GoldBio is excited to be the choice of researchers seeking to further understanding of the biology of crop plants important to populations throughout the world.

Category Code: 88241

Posted by Chris on March 6th, 2014  ⟩  0 comments

There is, I think, one thing that unifies nearly every species on Earth, a single constant that goes beyond similarities in DNA, genomes, cells or evolution. A “universal” tie-in which, even if you could genetically engineer and build from scratch an entirely new creation unbeholdened to any previous creature, rules every single body, great and small. Simply, we are all children of and are ruled by a planet with a 24 hour axis rotation.

This continuous cycle of light and dark creates the circadian rhythm that drives at the very core of our existence, mandating our waking and sleeping, whether human, insect, tree or bacteria. We live and die on the phases of our great home-world, and it takes thousands of years of isolation and/or specific adaption to overcome that basic, simple, biological drive. As humans, we often think that we have subjugated the world around us, but it’s abundantly clear to anyone who’s ever traveled long distances by plane (or even worked in a split-shift job) and suffered a “jetlag” effect, that we are still, quite obviously, slaves to our primal, internal clocks.

Despite our seeming dependence on it, we are still very much “in the dark” about how our circadian clock works. We know, for instance, that the clock is a function of the Suprachiasmatic nucleus (SCN), a small, rice-sized node in the hypothalamus that uses highly specialized retinal ganglion cells to detect light patterns, signaling genes such as Clock (Clk) and Period (Per) in the SCN to begin the oscillation of our internal clocks and maintain the speed of the oscillations over the next 24 hours. We know that there is an auto-regulatory feedback loop that prevents obtuse or episodic entrainment from occurring. But the way in which that regulation works hasn’t been as well understood...until just recently when a group out of Oxford led by Russell Foster and Stuart Peirson took an in depth look at the genes involved in the SCN feedback mechanism.

There is a real impetus to understand the way these genes process the external cues and self-regulate to govern our lives, and not just for jet-setting, world tourists, but also for the many people who suffer from circadian rhythm deficiencies. Foster and Peirson’s worked at the Per1 and Sik1 genes and how they affect CREB-mediated clock gene expression and that CRTC1 acts as a coactivator of the CREB transcription of those genes. They were able to show that knockdown of the Sik1 gene, both in vitro (with the smart use of Gold Bio Luciferin) and in vivo, enhanced the behavioral phase shifts and retrainment to new light-dark cycles by allowing the CRTC-continued transcription of Per1!

SCN Cirdadian Pathway

The role of the SIK1 negative-feedback mechanism is important to prevent our bodies from constantly readjusting to every minor change of stimuli that we’re faced with every day. Imagine if that streetlamp outside your window was suddenly able to change your body’s circadian clock to an opposite light-dark cycle or if, in the process of getting up in the middle of the night for a drink, the refrigerator light suddenly informed your body that it was morning and that you should stay awake? There is a definite need for the primary clock that we live by to keep itself in check, despite thousands of stimuli every day. That’s what the SIK1-CRTC pathway does…the proverbial time-keeping engineer on the train of life, keeping the system on time.

However there are always times when the clock goes haywire and needs a reboot. Currently, we don’t have one. But research into these mechanics and results like that of Foster and Peirson’s will someday make that a reality. And I can just about imagine the end to jetlag as we know it…but until then, I’ll just have to take a nap.

Jagannath, A., Butler, R., Godinho, S. I., Couch, Y., Brown, L. A., Vasudevan, S. R., Flanagan,K., Anthony, D., Churchill, G., Wood, M., Steiner, G, Ebeling, M., Hossbach, M., Wettstein, J., Duffield, G., Gatti, S., Hankins, M., Foster, R. & Peirson, S. (2013). The CRTC1-SIK1 Pathway Regulates Entrainment of the Circadian Clock. Cell, 154(5), 1100-1111.

Category Code: 79105 79102

Posted by Chris on February 27th, 2014  ⟩  0 comments

It is probably a safe bet that everyone reading this knows at least one person who has diabetes. Diabetes is such a common disease in our culture, we hardly blink anymore when we find out that someone has it, as opposed to more ominous diseases like cancer, cardio-vascular disease or AIDS. Diabetes is the common name for Diabetes mellitus, but it is also a generalized term that combines two very different diseases that produce similar symptoms and effects. When we hear the word “diabetes”, most immediately think of the Type 2 version, the overweight and unhealthy, dietary disease that affects nearly 10% of all Americans and more than 300 million people worldwide. That’s not surprising, since Type 2 diabetes (T2DM) makes up approximately 90% of all diabetes cases. But there’s a huge difference between Type 2 and Type 1 diabetes (T1DM).

Whereas T2DM can be largely traced to poor nutritional and lifestyle choices, along with a smattering of genetic predispositions, which collectively gang up against the body’s insulin and prevent it from doing its job, rather like the fairy tale Cinderella’s step-mother and sisters preventing her from going to the ball. However, T1DM is actually an autoimmune disease in which the body’s immune system goes haywire and begins targeting the insulin-producing beta cells in the pancreas. Imagine how bad that infamous story would have been if Cinderella’s had been killed by her wicked step-mother immediately after her father died instead.

T1DM isn’t just insulin resistance, it’s the loss of most or all of the insulin in the body, due to the destruction of the cells that produce it. The end-effect looks identical to T2DM, which is why they are both commonly called the same thing, but where Type 2 can be possibly be reversed with proper diet and better exercise, Type 1 is a lifelong condition that has little to no chance of ever going away. Also, to date, T1DM cannot be prevented. While it’s known what happens, we still do not know why; whether it’s a simply genetic susceptibility or some kind of external trigger or some combination of the two or even something else that’s yet to be discovered.

However, scientists have noticed patterns of autoantibodies that have been able to predict the onset of T1DM. Autoantibodies for islet cells (also called islet of Langerhans), the cells responsible for producing the beta cells which monitor sugar levels and release insulin, for insulin, for glutamic acid decarboxylase (GAD65), for protein tyrosine phosphatase (IA-2) and for zinc transporter 8 (ZnT8) all account for some degree of T1DM onset. The more, different autoantibodies found, the higher the likelihood of eventually developing T1DM. The presence of these autoantibodies is sometimes called “latent autoimmune diabetes” and there are some doctors who believe that if these autoantibodies can be detected and suppressed early enough, then full onset T1DM might be preventable.

Since insulin autoantibodies (IAA) are usually the first to appear, they are an enticing target for scientists and it should be an easy and simple ELISA experiment, but IAAs are additionally bothersome in that they refuse to bind to human insulin which has been bound to an ELISA plate. Several other methods, such as RIA (radioimmunoassay) and ECL (electrochemiluminescence), have therefore been developed. But RIA depends on radiolabeled antigens, which are always a pain to deal with, and ECL requires high cost equipment and training and there have been reports of poor result correlation between labs. But what if there was a way to make that ELISA work after all? Well, a group from France, led by Nathalie Morel, think they’ve found a way.

Bridge ELISA

Morel’s concept is called a Bridge-ELISA. Binding the IAA to a biotinylated insulin in one of its antigen-binding domain (for a great system that can do just that, check out our Gold Bio Biotin Labeling Kits) allowed for easy detection via a streptavidin-conjugated tracer, and the other of its antigen-binding domains was bound to a GC300 hapten to help bind to an ELISA plate coated with the anti-GC300 monoclonal antibody MC159. Ultimately, Morel’s group developed a process that is faster, simpler to use and more reliable than ECL, and was safer (since there is no radioactive components) than the traditional RIA method, although it was only about 80% as sensitive as RIA in detecting IAA’s in T1DM children. Because it utilizes the standard ELISA format, practically every protein lab in the world can do it and automate the process for large-scale, faster results. And when it comes to saving insulin producing beta cells and possibly preventing T1DM, every small step forward might help to save a life you know.


Kikkas I, Mallone R, Tubiana-Rufi N, Chevenne D, Carel JC, Creminon, C., Volland, H., Boitard, C. and Morel, N. (2013) A Simple and Fast Non-Radioactive Bridging Immunoassay for Insulin Autoantibodies. PLoS ONE 8(7): e69021. doi:10.1371/journal.pone.0069021

Category Code: 88241 88231