Dear all,
I would like to transfect protoplasts with constructs made using the MoClo technique, but it seems that the vectors in my toolkit are specifically made for Agrobacterium-mediated transfer. Are there any vectors available for protoplast transfection?
Thank you in advance!
Kind regards, Jonas De Saeger
Dear Jonas De Saeger
Pls. find the attached files
http://2016.igem.org/Resources/Plant_Synthetic_Biology/Plants_and_iGEM
http://pubs.acs.org/doi/full/10.1021/acssynbio.6b00082
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4757859/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4730745/
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0016765
http://pubs.acs.org/doi/abs/10.1021/acssynbio.6b00089?journalCode=asbcd6
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC241498/
http://www.mdpi.com/1422-0067/17/1
http://www.microbiologyresearch.org/docserver/fulltext/micro/133/3/mic-133-3-493.pdf?expires=1481734452&id=id&accname=guest&checksum=9B13A1F0A78342916334D918B48778BA
http://worldwidescience.org/topicpages/p/progenitor+cells+transfected.html
page 219. In: A.T. Ganesan. 2012. Bacillus Molecular Genetics and Biotechnology Applications Academic Press,Pp.497. ISBN 0323138632, 9780323138635
http://onlinelibrary.wiley.com/doi/10.1111/j.1574-6968.1987.tb02467.x/abstract?systemMessage=Wiley+Online+Library+will+be+unavailable+on+Saturday+17th+December+2016+at+09%3A00+GMT%2F+04%3A00+EST%2F+17%3A00+SGT+for+4hrs+due+to+essential+maintenance.Apologies+for+the+inconvenience
Hoping this will be helpful
Regards
Prof. Houda Kawas
In principle there is nothing specific about a vecctor used for protoplast transformation. A vector designed for biolistics is also suitable for protoplast work. That is a vector in the size range of 6-7 kbp rather than a binary vector around 12 kbp is preferable. If you have already constructed your binary vector, you could just use restriction enzymes to cut out the DNA, purify that DNA and use that to transfect your protoplasts.
Can you try the following kit
MoClo Plant Parts Kit (deposited by Nicola Patron's lab )
- Contains standardized parts specific for plant synthetic biology
Please check this link below,
https://www.addgene.org/cloning/moclo/
Thank you all for the answers!
@ Vidhu Sankar Babu: this is indeed the kit we use, but as far as I know all of the vectors in this toolkit are designed for use in Agrobacterium?
@ Gregory D Nugent: thank you for your reply; in other words, I can also use binary plasmids for protoplast transfection (keeping into account that they are quite large)?
@ Houda Kawas: thank you for the wide variety of sources.
Kind regards, Jonas
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