Construction of Chloroplast Transformation System

Chloroplast transformation systemsare based on homologous recombination to insert foreign DNA into specific chloroplast genomic loci. As early as the 1990s, people obtained stable chloroplast transgenic plants of tobacco. In recent years, tomato, lettuce, rape, potato, cabbage, cotton and carrot have also been used as target plants for plastid transformation technology, which revolutionized the study of gene expression regulation mechanism. In addition, researchers focus on the research of chloroplast genetic transformation technology with metabolic pathways, biological agents, vaccines, etc. At present, plastid transformation technology has become an important method to analyze the function of chloroplast genes, and is also widely used in the mass production of exogenous proteins for agronomic traits.

Fig. 1. Diagrammatic representation of the processes for chloroplast genome transformation. (Chen P Y, et al., 2020)

Services

The primary goal of establishing chloroplast transformation technology is to construct various chloroplast expression vectors for different plant species, and then introduce the constructed expression vectors into the chloroplast genome to obtain the desired chloroplast transformed plants. Over the years, the team at Lifeasible has accumulated rich experience in chloroplast genetic engineering and has built a mature chloroplast genetic engineering platform. Our scientists are committed to providing chloroplast transformation system construction services to constructe specific chloroplast expression vectors for various plants (such as spinach, lettuce, tomato, bitter gourd, microalgae, tobacco), and evaluate the function of the constructed vectors. We provide the following strategies to achieve the construction of chloroplast transformation system:

• Particle Bombardment-Mediated Chloroplast Transformation
• Polyethylene Glycol (PEG)-Mediated Chloroplast Transformation
• Agrobacterium-Mediated Chloroplast Transformation

In addition, we introduced the constructed specific expression vector into the chloroplast genome based on the method of chloroplast gene transformation, and use various methods to detect whether the foreign gene is integrated into the chloroplast genome for expression. Based on the following plastid transformation process, we can provide customers with high-quality chloroplast transformation system construction services.

(1) PCR amplification of chloroplast homologous sequences.
(2) Synthesize exogenous expression cassettes by chemical synthesis.
(3) Obtaining a chloroplast transformation vector.
(4) The transformation vector carries the exogenous target gene and is introduced into the chloroplast by biolistic method or other transformation systems.
(5) Integrating the exogenous expression framework into the chloroplast genome.
(6) Screening of transformed chloroplast cells.
(7) Subculture to obtain stable chloroplast transformed plants.

Methods for Constructing Chloroplast Transformation System

According to the main process of chloroplast transformation, we have successfully developed and established a chloroplast gene transformation platform, adopting a variety of chloroplast gene transformation methods to accelerate your research.

• Microbomb bombardment.
• Agrobacterium-mediated transformation methods.
• PEG treatment.
• Microinjection.
• Carbon nanotube carrier.
• Ultraviolet laser microbeam.

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. Chen P Y, Tsai Y T, To K Y. (2020) Construction and Evaluation of Chloroplast Expression Vectors in Higher Plants[M]//Genetic Transformation in Crops. IntechOpen.
2. Narra M, Kota S, Velivela Y, et al. (2018) Construction of chloroplast transformation vector and its functional evaluation in Momordica charantia L[J]. 3 Biotech. 8(3): 1-11.
3. Adem M, Beyene D, Feyissa T. (2017) Recent achievements obtained by chloroplast transformation[J]. Plant Methods. 13(1): 1-11.