Overview of Quality Assessment for Genetically Engineered Strains
Microbes are everywhere and play vital roles in various industries, from agriculture to medicine and biotech innovations. While beneficial microbes can significantly impact your projects, there's a risk if you don't thoroughly understand what you're working with. Contamination can wreck products, lead to failed experiments, or even create unsafe conditions-none of which are ideal outcomes. That's why we initiated the Microbial Strain Identification and Quality Assessment Service, aiming to provide clear and accurate data on the microbes in your work environment.
We don't promise miracles; we deliver reliable results and straightforward testing that genuinely make a difference. Our service focuses on confirming that your microbes are exactly what you believe them to be-pure, safe, and ready to perform. We utilize high-quality tools and bring the expertise to match, but keep our approach practical and clear. We believe that everyone, whether you're a small startup or a big corporation, deserves to know exactly what's in their samples. With our support, you'll gain peace of mind, allowing you to focus on what truly matters and trust that your microbes are up to the task.
Common Methods for Quality Assessment of Genetically Engineered Strains
PCR (Polymerase Chain Reaction)
PCR is a go-to technique for spotting specific DNA sequences in modified strains. It's quick and can zero in on target genes, like marker or reporter genes used in modifications. Its high sensitivity makes it perfect for checking genetic tweaks and keeping purity in check.
MALDI-TOF MS (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry)
MALDI-TOF MS excels at pinpointing microbial strains fast and accurately. By comparing protein profiles of unknowns to a known database, it nails down strain identities. It's super accurate—like over 97% for yeast strains—and great for confirming modified strains and spotting unintended changes.
Genetic Transformation Techniques
Methods like electroporation, biolistic bombardment, and Agrobacterium-mediated transformation introduce new genes into strains. Electroporation is super-efficient, especially for nuclear transformations in microalgae. These techniques are key for creating engineered strains, often checked with PCR or sequencing for success.
Genome Editing Tools (CRISPR/Cas9, TALENs)
Tools like CRISPR/Cas9, TALENs offer precision edits in microorganism DNA. CRISPR/Cas9 has transformed genetic engineering, enabling precise tweaks with great accuracy. These tools are vital for crafting and validating strains to ensure they meet specific genetic goals.
Microbial Contamination Analysis
It's crucial to keep engineered strains free from contaminants. PCR and MALDI-TOF MS help screen for any unwanted guests, ensuring your strains are both pure and stable. This step is essential for upholding genetic integrity and ensuring strains are safe and effective for research or industrial use.
Applications and Industries Served
Biotechnology and Pharmaceuticals
In biotech and pharma, quality checks for engineered strains are crucial. These sectors use modified strains for producing things like therapeutic proteins and biofuels. Engineered E. coli, for example, is vital for insulin production. Ensuring these strains are pure and stable is key to product safety.
Agriculture
In agriculture, engineered strains enhance crop resilience and yields. Genetic tweaks enable crops to resist pests, diseases, and environmental stress. CRISPR/Cas9, for instance, helps create crops with better fungal resistance, reducing pesticide use. Quality checks ensure these strains remain effective, supporting sustainable farming and food security.
Research Institutions
Quality assessment is critical for reliable research. Scientists use engineered strains to study biology and innovate. Our service offers genetic analysis and contamination screening, ensuring research is based on stable, pure strains. This support is vital for achieving accurate results and high-quality publications.
Regulatory Compliance
Meeting regulatory standards is essential when using genetically modified strains. Rigorous testing is required to ensure safety for humans and the environment. Our service provides comprehensive reports confirming genetic purity and stability, helping products meet the necessary approval standards.
Our lab is equipped with state-of-the-art tools for microbial strain identification and quality checks. We have the DW-M80 for fast and precise biochemical and genetic tests. For protein profiling, we use MALDI-TOF MS, which provides quick and accurate results. Our real-time PCR equipment is essential for DNA detection, while HPLC helps us analyze metabolites in detail. For genomic studies, we rely on next-generation sequencing technology. Additionally, we have automated systems for large-scale cultivation, setups for antigen capture, and microarrays for high-throughput screening. These advanced instruments ensure that we can provide comprehensive and reliable quality assessment for genetically engineered strains.
Expertise and Experience: Our team has deep experience in microbial physiology and genetic engineering, ensuring dependable assessments of engineered strains.
Comprehensive and Accurate Results: We offer detailed evaluations that cover genetic purity, stability, and contamination, giving you a full understanding of your strains.
Customized Solutions: We shape our services to match your needs in biotech, agriculture, or research, offering tailored approaches.
State-of-the-Art Technology: We use cutting-edge tools like PCR, sequencing, and MALDI-TOF MS for precise and accurate analysis.
Case Study
Case Study 1: Antibiotic-Free Cloning Method Achieves Efficient Plasmid Stability
Researchers set up a pipeline for cloning without antibiotic resistance genes, using a strategy that relies on auxotrophic and essential gene selection. It works by constructing plasmids in specially modified E. coli DH10B strains and then using both selection methods to pick out recombinant strains and keep plasmids stable in E. coli Nissle 1917 and E. coli MG1655. These are standard strains used in engineered probiotics and lab experiments, respectively. This method matches the efficiency of traditional antibiotic resistance cloning. Plus, the double-knockout Nissle and MG1655 strains are easy to work with. Importantly, engineered Nissle strains can maintain plasmids over time, even in mouse guts, suggesting this system could be broadly useful while reducing the risk of spreading antibiotic resistance.
Fig. 2. Transformation efficiency of auxotrophic strains with in vitro Gibson-assembled low-, medium, and high-copy plasmids containing thyA or glmS expression cassettes. (Amrofell, et al., 2023)
Case Study 2: Standard plate counts to detect Listeria. PCR to confirm and check antibiotic resistance genes.
Researchers looked at Listeria monocytogenes, a nasty foodborne germ that can spread through soil and water, causing serious illness. They tested water and soil samples from two districts in South Africa to see how common it was and how well it resisted antibiotics. Researchers found the germ in both water and soil, with some samples having pretty high levels. All the samples had genes that make them dangerous. Most were resistant to several antibiotics, with tetracycline and doxycycline being the most common. Some even had genes for extra-strong resistance. This means the water and soil could be spreading drug-resistant Listeria into the food supply, which is a big concern for public health.
Fig. 3. Heatmap showing the phenotypic antibiotic resistance patterns of each L. monocytogenes isolates. (Iwu, et al., 2020)
Case Study 3: Study Identifies Toxin-Producing Fungi in Soil Invertebrates in Taif, Saudi Arabia
Harmful aflatoxins, caused mostly by Aspergillus fungi, are a global health issue. This study in Taif, Saudi Arabia, identified fungi from soil invertebrates, focusing on toxin production. Out of 114 fungal samples, 22 were Aspergillus. Using gene sequencing and PCR, researchers grouped these fungi into clusters and confirmed some produce toxins like aflatoxin B1 and ochratoxins.
Fig. 4. Morphological characteristics of Aspergillus species isolated from soil invertebrates in Taif, Saudi Arabia. (Awad, et al., 2023)
FAQs
Q: What samples can you analyze?
A: We can handle a range of samples like bacterial and fungal strains, environmental samples, food products, and fermentation samples from industries.
Q: Can you find antibiotic resistance genes?
A: Yes, we use PCR and advanced techniques to screen for antibiotic resistance genes.
Q: What reports will I get?
A: You'll get a detailed report with ID results, sequence data, phylogenetic analysis, plus any found virulence or resistance genes.
Q: Do you analyze probiotics or food additives?
A: Yes, we offer full analysis for probiotics, food additives, and other microbial products in the food industry.
References:
Iwu CD, Okoh AI. Characterization of antibiogram fingerprints in Listeria monocytogenes recovered from irrigation water and agricultural soil samples. PLoS One. 2020;15(2):e0228956.
Awad MF.; et al. Identification and biodiversity patterns of Aspergillus species isolated from some soil invertebrates at high altitude using morphological characteristics and phylogenetic analyses. PeerJ. 2023;11:e15035.
Amrofell MB.; et al. Engineering E. coli strains using antibiotic-resistance-gene-free plasmids. Cell Rep Methods. 2023;3(12):100669.
We are here to help you further your development in the microbiology field.
Fig. 2. Transformation efficiency of auxotrophic strains with in vitro Gibson-assembled low-, medium, and high-copy plasmids containing thyA or glmS expression cassettes. (Amrofell, et al., 2023)
Fig. 3. Heatmap showing the phenotypic antibiotic resistance patterns of each L. monocytogenes isolates. (Iwu, et al., 2020)
Fig. 4. Morphological characteristics of Aspergillus species isolated from soil invertebrates in Taif, Saudi Arabia. (Awad, et al., 2023)
