Engineering Bacterial Operons by Chloroplast Genomes

As a non-autonomous organelle, the chloroplast genome moved toward the nucleus during evolution but retained its unique genomic features, such as circular genome structure, gene organization in bacterial-like operon transcriptional units, etc. An operon is formed by DNA units composed of multiple genes that often share a common function under the control of a single promoter. Scientists have succeeded in predicting bacterial operons by utilizing a supervised machine learning algorithm trained on experimental data. But few analyses of chloroplast operons have made the fundamental elements of these structures unclear, limiting their ability to use them in synthetic biology. Scientists have now used the Bt cry 2Aa2 operon as a model system to test the feasibility of expressing polygenic operons in engineered chloroplasts.

Fig. 1. Creating a generic dataset of plastid operons composed of diverse evolved organisms. (Shahar N, et al., 2019)

Services

The ability to convert synthetic genes into plastids is currently limited by the lack of large-scale information on chloroplast operons. In addition, inferring the operon map of the chloroplast is helpful for understanding its genetic regulation and also for gene annotation. Over the years, the team at Lifeasible has accumulated extensive experience in chloroplast genetic engineering and is commmitted to engineering bacterial operons through the chloroplast genome. Here, we provide complete and reliable chloroplast primary operon prediction and large-scale analysis services to help you understand unique genetic features in chloroplast evolution, which supporting your innovative discoveries in chloroplast genetic engineering.

Our engineers combined experimental tools and predictive modeling to successfully create a database of chloroplast-specific operons. And a complete protocol has been developed to analyze the entire operon map of higher plant chloroplasts, including the composition and characteristics of chloroplast operons.

(1) Construction of chloroplast operon marker dataset.
(2) Creating an energy spectrum of RNA folding for each gene.
(3) Performing a wrapped reverse elimination feature selection on the entire dataset. And select a high-precision feature set with a small number of features.
(4) Creating a chloroplast operon prediction model and evaluate the performance of the model.
(5) Analysis of plastid operon signatures by analyzing this newly formed database.

Advantages of Our Services

• Realizing the use of the natural ability of chloroplasts to express polycistrons and design vectors with multiple genes under the control of a single promoter.
• Selecting operons of higher plants, highly reliable and contain all genes.
• Ability to highlight essential features that distinguish operon pairs from non-operon pairs.
• Ability to grasp the main fundamental features specific to operon genes and avoid potential species-specific biases.
• Minimizing plasmid size and allowing the introduction of several metabolically relevant transgenes in a single transformation.
• Providing support for the advancement of synthetic biology and basic science (metabolic engineering) in the chloroplast.

Lifeasible can meet the needs of customers on time and on budget through a wide range of chloroplast genetic engineering strategies. Our aim is to be customer-centric and to provide the highest quality service to customers around the world. Our skilled and dedicated scientific researchers ensure that the most appropriate methods and techniques are selected for each specialized chloroplast project. Our customer service representatives are enthusiastic and trustworthy 24 hours a day, 7 days a week. If you are interested in our services, please feel free to contact us for more information or a detailed discussion.

References

1. Daniell H, Khan M S, Allison L. (2002) Milestones in chloroplast genetic engineering: an environmentally friendly era in biotechnology[J]. Trends in plant science. 7(2): 84-91.
2. Shahar N, Weiner I, Stotsky L, et al. (2019) Prediction and large-scale analysis of primary operons in plastids reveals unique genetic features in the evolution of chloroplasts[J]. Nucleic acids research. 47(7): 3344-3352.