How efficient is Agrobacterium in expressing foreign proteins from non-plant members such as algae and cyanobacteria in potato chloroplast? Can you please suggest me some reliable protocols if any?
Hi Sarbesh, are you asking about transforming the potato chloroplast genome itself, or about transforming the potato nuclear genome and expressing the proteins in the chloroplast?
your question is not clear enough .... do you mean transferred plasmid role on Rubicon synthesis participation between chloroplasts and nucleus..... or what
Thank you for your kind interest, sir. I am asking about genetic engineering at the chloroplast genome level and expression of foreign protein at the chloroplast itself and not at the nuclear genome level.
Using Agrobacterium mediated transformation in potato , You can direct the expression of foreign gene in chloroplast by using transit peptide. It will be genetic transformatin however your gene will get expressed in chloroplast. Ypu can use RubisCo gene promote/ or engineer TP region upstream to your gene.
If you are talking about transforming the chloroplast genome itself (the plastome) this is generally done by particle-mediated transformation (although there have been some extremely clever results from Japanese Riken group using cell penetrating peptides complexed with DNA and transit peptides as well). I don't know of any reliable method of using Agrobacterium to transform the chloroplast genome.
If you are looking to just express the introduced protein in the chloroplast, transforming the nuclear genome with Agrobacterium and using a transit peptide might be the most efficient method, as Allah Bakhsh suggests.
1. Plastid transformation is a good tool for high-level 'recombinant protein' production due to numerous copies of chloroplasts in a cell. It also reduce the environment concern due to (trans)gene flow through pollens because they are contained in the cytoplasm.
2. So far, the most efficient plant for plastid transformation is tobacco. Many papers have published plastid transformation using tobacco. However, with a good protocol you should be able to see a reasonable efficient plastid transformation for potato. See this paper (2014) for a improved protocol for potato plastid transformation.
http://www.ncbi.nlm.nih.gov/pubmed/24599861
Methods Mol Biol. 2014
Plastid transformation in potato: Solanum tuberosum..
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Microorganisms
- Bacteria