As the global scientific community celebrates the 70th anniversary of the discovery of DNA, 2023 also marks the anniversary of the powerful technology that revolutionized PCR 20 years ago. In 2003, the introduction of Thermo Scientific™ Phusion™ High-Fidelity DNA Polymerase enabled high-performance PCR and ushered in an era of continuous advances in fusion protein technology designed for superior fidelity of DNA replication.
Each Phusion product introduction—from Thermo Scientific™ Phusion™ Hot-Start Polymerase to the latest Thermo Scientific™ Phusion™ Plus DNA Polymerase—has provided researchers with new applications that leverage the excellent fidelity, speed, specificity, and sensitivity of the enzyme. Over the years, the variety of Phusion master mixes and kits optimized for different applications have helped researchers explore new possibilities in fields such as genetics, medicine, and biotechnology. In celebrating fusion protein technology, we’ve compiled some of the most unique and fascinating research projects published in recent years using this next-generation PCR technology.
1. Cleaning up oil spills
Most petroleum spills occur on land, not water, but the ecological and health impacts are devastating wherever they occur. Researchers have been investigating the use of plants to remediate sites contaminated with petroleum. In a recent study, scientists used Thermo Scientific™ Phusion™ High-Fidelity DNA Polymerase to help study the genes of bacteria species capable of promoting the growth of plants resistant to diesel fuel hydrocarbons. The successful results of the study point to a future where bacteria and plants may be deployed to work together synergistically and ecologically to clean up fuel spills.
2. Improving orthodontic healthcare
Acrylic retainers worn after orthodontic procedures to help align teeth must be cleaned to prevent the development of biofilms on the surfaces of the retainers. In a unique study, investigators used PCR amplification with Phusion Plus DNA Polymerase to examine oral microbiota changes from the wearing of retainers to help evaluate the effectiveness of a new chemical cleaning product. The study found that the most effective method of eliminating retainer biofilms involved a combination of both chemical and mechanical cleaning (brushing).
3. Creating industrial “cell factories”
The yeast Saccharomyces cerevisiae is widely used in industrial biotechnology to produce fuels, chemicals, food ingredients, and pharmaceuticals. However, obtaining high‐performing strains for such bioprocesses often requires extensive and costly testing of metabolic engineering targets. Research at the Technical University of Denmark using CRISPR Cas9 gene editing and PCR amplification with Phusion DNA polymerases led to a method that allows for simultaneous manipulation of several metabolic engineering targets. This method helps accelerate efforts to construct cell factories for industrial uses.
4. Improving agricultural biopesticides
Insects are remarkably tenacious—and can be equally destructive. Bt toxin-producing crops have been widely adopted for agricultural pest management, leading to considerable economic and environmental benefits. However, Bt toxin resistance has evolved among insect pests, posing a serious threat to this pest control strategy. In a study published in Nature, researchers developed, using Thermo Scientific™ Phusion™ U Hot Start DNA Polymerase, a system to evolve Bt toxin variants with novel insect cell receptor affinity that can overcome Bt toxin resistance in insects.
5. Tackling tick-borne diseases
If you are familiar with lyme disease, you know that tick-borne infections are a serious threat to humans. Tick-borne infections also pose a threat to livestock and companion animals and can lead to animal illness and death. Molecular identification of pathogens within the tick vectors that cause these infections can be challenging because ticks sampled from animals are often engorged with animal blood, which can have an inhibitory effect on PCR enzymes, making it difficult to amplify nucleic acid. Researchers comparing different methods of DNA extraction from blood-engorged ticks found that Phusion Plus PCR Master Mix was the best approach for amplifying the tick 16s rRNA gene, regardless of extraction method.
6. Building biodegradable polymers with the help of slime proteins
While they pose no threat to humans, velvet worms (Onychophora) attack their prey (such as termites and small spiders) with a powerful weapon: sticky slime they shoot with precision from up to a foot away. Upon ejection, the sticky goo envelops the prey and undergoes a rapid liquid-to-solid transition. The slime is a strong and fully biodegradable protein material, which could inspire the sustainable fabrication of useful polymers. Combining transcriptomic and proteomic studies, Researchers used Phusion Plus Polymerase to verify the sequences of slime proteins, obtaining the complete primary sequences of slime proteins, and identifying key features of slime self-assembly. The study opens a research path for the development of biodegradable industrial polymers.
Find the right Phusion high-fidelity DNA polymerase format for your application at thermofisher.com/phusion
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- Eze, Michael O., et al. Bacteria-plant interactions synergistically enhance biodegradation of diesel fuel hydrocarbons. Communications Earth & Environment 3.1 (2022): 192.
- Ref. Kasibut, Punnisa, et al. Oral Microbiome in Orthodontic Acrylic Retainer. Polymers17 (2022): 3583.
- Kildegaard, Kanchana Rueksomtawin, et al. CRISPR/Cas9‐RNA interference system for combinatorial metabolic engineering of Saccharomyces cerevisiae. Yeast5 (2019): 237–247
- Badran, Ahmed H., et al. Continuous evolution of B. thuringiensis toxins overcomes insect resistance. Nature 533(7601) (2016): 58–63.
- Reifenberger, Gretchen C., Bryce A. Thomas, and DeLacy VL Rhodes. Comparison of DNA Extraction and Amplification Techniques for Use with Engorged Hard-Bodied Ticks. Microorganisms6 (2022): 1254.
- Lu, Yang, et al. Complete sequences of the velvet worm slime proteins reveal that slime formation is enabled by disulfide bonds and intrinsically disordered regions. Advanced Science18 (2022): 2201444.
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