Optimization of Foreign Gene Expression in Chloroplast

After long-term in-depth research on the structure and expression of chloroplast genes, people are familiar with the complete chloroplast genome sequences of various plants, and have accumulated a large amount of DNA sequence data. Since chloroplast gene transformation engineering can explore the mechanism of transgene expression in vivo, it has attracted much attention. This technology is mainly to transfer exogenous DNA into the chloroplast of higher plants through a variety of advanced methods for transformation. Therefore, foreign genes play a key role in this technology. Many studies have reported that levels of exogenous protein produced in transgenic chloroplasts exceed 10% of the total plant soluble protein. However, transgenes transformed by the chloroplast genome may have low or no expression in the chloroplast. Scientists have engineered additional translation-enhancing elements from non-plant sequences to enhance the expression of exogenous genes in the chloroplast. Optimizing the process of delivering exogenous DNA to the chloroplast of higher plants has become a research hotspot.

Fig. 1. Construction of chloroplast vectors using native or codon-optimized genes, and evaluation of homoplasmy and transgene expression. (Kwon K C, et al., 2016)

Services

Over the years, the team at Lifeasible has accumulated extensive experience in chloroplast genetic engineering. Here, we provide professional and reliable foreign gene expression optimization services in chloroplasts to achieve high-level expression of single transgenes in higher plant chloroplasts. Our engineers specialize in building a set of feasible exogenous gene incorporation or replacement systems to help customers conduct in vivo studies of chloroplast gene expression, regulation, identification of unknown chloroplast genes, and open reading frames. In addition, we employ a variety of methods to regulate genes in the chloroplast, including the generation of chloroplast mutants, protoplast fusion, organelle inactivation, chloroplast recombination, etc.

Our engineers rigorously considered the multiple factors that determine the efficiency of plastid transgene expression and developed appropriate strategies to maximize transgene expression. Our results will guarantee your 100% satisfaction.

· Choice of promoter.
· Choice of 5'-UTR.
· Choice of 3'-UTR.
· Genomic integration site.
· Codon usage.
· RNA secondary structure.
· Protein stability.
· Protein localization.
· Choice of host plant.
· Choice of growth conditions.
· Choice of plant tissue types.
· Choice of developmental stages of plant material.

Advantages of Our Services

• An efficient and highly repeatable delivery system.
• Considering multiple factors affecting chloroplast transformation, not only the 5'UTR element, but also the sequence downstream of the initiation codon was optimized.
• Accelerates the understanding of DNA replication, recombination, transcription, translation and protein import in the chloroplast by regulating the expression of foreign genes.
• Levels of exogenous proteins can be manipulated over an extraordinary 10,000-fold range.
• Transformed chloroplasts can be reintroduced into protoplasts.

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. Kwon K C, Chan H T, León I R, et al. (2016) Codon optimization to enhance expression yields insights into chloroplast translation[J]. Plant physiology. 172(1): 62-77.
2. Ye G N, Daniell H, Sanford J C. (1990) Optimization of delivery of foreign DNA into higher-plant chloroplasts[J]. Plant molecular biology. 15(6): 809-819.