Expression of Exogenous Proteins in Chloroplasts

Expression of Exogenous Proteins in Chloroplasts

Plant chloroplasts diverged from preplastids and are evolutionarily derived from endosymbiotic bacteria. As a result, chloroplasts retain many of the characteristics of prokaryotic cells, such as multiple cis-trans, gene transcription, and protein translation, all of which are similar to those of bacteria. However, over millions of years of evolution, most of the chloroplast genes have been transferred to the nuclear genome, which retains three main categories of functional genes; photosynthesis, expression regulation, and biosynthesis. Plant genetic engineering techniques have received increasing attention to enable the use of plant chloroplasts as bioreactors for synthesizing exogenous proteins.

Figure 1. Overview diagram of chloroplast transcription.Figure 1. Overview diagram of chloroplast transcription. (Yagi, Y.; Shiina, T. 2017)

Lifeasible has been working in the field of plant genetic engineering technology for many years, and we have considerable success in using plant chloroplasts as bioreactors for biopharmaceuticals, expression of pharmaceutical proteins, and vaccine biosynthesis. We are confident that we can provide you with the right solution to enable the expression of your target exogenous protein in plant chloroplasts.

What We Offer

We first construct the homologous recombinant fragment, the expression cassette of the selection marker gene, the expression cassette of the exogenous target gene, and its promoter and terminator into the same expression vector. After constructing the expression vector, the vector is introduced into the chloroplast using an efficient transformation method. After homologous recombination, the exogenous gene is expressed as the chloroplast genome is replicated and inherited.

Methods of Chloroplast Transformation

Currently, the genetic transformation of chloroplasts is generally carried out by introducing transgenes into the chloroplast genome by the gene gun method, glass bead transformation, electroshock transformation, or PEG-mediated transformation. In addition, we can use lipid exchange membrane permeation (LEP) technology to transport genes into the chloroplasts of plant cells without external forces with the aid of nanoparticles to transport DNA.

Advantages of Synthesizing Exogenous Proteins in Plant Chloroplasts

  • Chloroplast DNA is inherited through the maternal line, and there is little risk of undesirable spread of the transgene.
  • Many chloroplast copies are present in each plant cell, allowing for high expression of exogenous genes.
  • Proteins expressed in chloroplasts can form disulfide bonds, and human-derived proteins can be folded correctly when expressed.
  • Integrating of exogenous genes into the chloroplast genome by homologous recombination does not produce positional effects or gene silencing.
  • Most chloroplast genes exist as manipulates, allowing for the simultaneous transformation and expression of multiple exogenous genes in the chloroplast genome.

Plant Species Covered by Our Services

  • Nicotiana tabacum
  • Arabidopsis thaliana
  • Solanum tuberosum
  • S.lycopersicum
  • Lesquerellafendleri
  • Petunia hybrida
  • Chlamydomonas reinhardtii
  • Glycinemax L.
  • Daucuscarota L.
  • Gossypium hirsutum
  • Lactucasativa L.
  • Oryzasativa, etc.

Exogenous Proteins That Have Been Successfully Expressed in Plant Chloroplasts

  • Human therapeutic protein
  • Aminoglycoside adenylyases
  • Renal luciferase
  • Green fluorescent protein (GFP)
  • Foot-and-mouth disease virus VP1 antigen protein
  • Human soluble TRAIL protein
  • Human interleukin (hIL4)
  • Human glutamic acid decarboxylase (hGAD65)
  • Rabies virus vaccine
  • pfs25 and pfs28 antigenic proteins etc.

Applications

  • Production of edible protein
  • Improvement of agronomic traits
  • Production of vaccine proteins
  • Production of antibody proteins
  • Production of critical pharmaceutical proteins, etc.

Our Service Process

Our Service Process

With outstanding experts and abundant plant genetic engineering technologies, Lifeasible provides customers in the field of biosynthesis with solutions for expressing exogenous proteins using plant chloroplasts. We sincerely look forward to cooperating with you; please feel free to contact us if you have any questions.

Reference

  1. Yagi, Y.; Shiina, T. (2014). Recent advances in the study of chloroplast gene expression and its evolution. Frontiers in plant science, 5, 61.
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