Can all bacterial plasmids work in plants too? - (Aug/29/2006 )
hi, i recently got a new plasmid...pQBI T7-GFP from a friend in my lab. She is using it just in e.coli but i was hoping to transform some of my plants with it using a gene gun. I have a sequence map of the plasmid, is there any reason why the GFP gene wouldnt work in a plant? It also codes for ampicillin resistance, does anyone know about ampicillin and plant toxicity? How much of it should I add to the plant growth media...I cant seem to find much data on this in google. thanks a lot.
mmmmmmmmh so many interesting, stimulating questions!!! (today I am a bit of a nerd! )
Let's start somewhere...
I don't know about that particular plasmid, but there are many things you have to check before using it in your plants...for example: what promoter controls the gfp-gene? is it constitutive or regulated? in that case: can you induce it in planta without risks for your tissues? (e.g.: if you have to use a chemical inducer, it may be toxic for your plant...)
About ampicillin: it's quite common, but its effect will be strongly dependent on your plant species, explant, growth condition etc. etc. etc. If you do not have any previous data set, you have to perform a preliminary toxicity assay using the growth conditions that you will establish for your experiment. Usually it's time-consuming, but it's also necessary...
hope this helps!
All bacterial plasmids don't even work in all bacteria -- thus the set of plasmids known as suicide vectors. Additionally, not all genes on bacterial plasmids work in all bacteria. For example, I routinely use a plasmid that replicates in E. coli, but can not replicate in Bacteroides fragilis. This plasmid also confers gentamicin resistance when in B. fragilis, but not when in E. coli (where it confers ampicillin resistance).
We'll clone DNA into this plasmid using E. coli as the host, and introduce it into B. fragilis, and select for gentamicin resistance, thus selecting for integration of the plasmid directed by homologous recombination, and achieving the first step in allelic exchange.
We have plasmids that work in B. fragilis, but not in B. distasonis...
Whether any given bacterial plasmid would work (replicate and/or express plasmid borne genes) when jumping from Bacteria to Eukarya is unknown. Certainly there are examples (the plasmids of Agrobacterium, Rhizobium, and Bacillus species spring to mind), but it would be a long shot, unless the plasmid was specifically engineered to be a chimera shuttle plasmid, and thus is able to replicate in both.
i think there is T7 promoter in that plasmid , hence wont really express in plant since plants need 35S promoter or others , nless its in a plant viral vector ,,,, then the process is different , but u wont get it expressed in the next generation only in that infected one
but then again if u try cloning it into a plant expression vector and introduce it into agrobacterium , and then transform the plant it might work ..
there are shuttle vectors like pPZP212 vectors that can work both in ecoli and agrobacterium and got the TDNA region that will transfer the GFP gene to the plant ( work already been done unfortunately ) a
also check the gfp gene sequnece
in a previous work they reported that a cryptic intron in the N terminal of GFP gene wild type didnt produce much flouresence , hence they modified it to get it more expressed in plants
if ur intrested i can send u that paper
I am intrested too. Could you kindly send me that paper or give me the bibliographic reference?
Thank you very much in advance!
i am sending the gfp modification paper
Thank you very much!