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Project using Agrobacterium tumefaciens

Posted: Fri Jul 12, 2019 4:05 am
by deleted-738009
Hey!
I am doing a project on the genetic modification of soybeans (to transform the soybean to be immune to herbicides) using Agrobacterium tumefaciens. I was wondering whether I needed to grow the Agrobacterium tumefaciens or if i could buy it :D somewhere?
Thanks so much!

Re: Project using Agrobacterium tumefaciens

Posted: Fri Jul 19, 2019 12:43 pm
by deleted-674917
Hi Kristen,

I found at least two inexpensive sources for buying Agrobacterium.
https://www.carolina.com/bacteria/agrob ... /154825.pr and
https://www.wardsci.com/store/product/8 ... umafaciens

There are also multiple companies selling competent cells of Agrobacterium which can take up your DNA of choice for infection.

Hope this helps!
MS

Re: Project using Agrobacterium tumefaciens

Posted: Tue Jul 23, 2019 4:59 am
by deleted-738009
Thank you very much for your reply!

Sorry I should have said that I live in australia and I can't find any stock here and i don't think you can ship bacteria overseas.

For my project I was hoping to grow a round-up ready soybean crop (tolerant to glyphosate) however I am not sure what the method would be to genetically engineer the agrobacterium tumefaceins so that it would insert the round up ready gene into the soybean (I also am not sure what the would be and where to get this gene from). Please please can you help :D

Re: Project using Agrobacterium tumefaciens

Posted: Tue Jul 23, 2019 2:21 pm
by deleted-674917
The best option I think is to get in touch with a lab (preferably in Australia or elsewhere) that performs research using Agrobacterium and request them for the reagents you need. Most academic labs are usually willing to share reagents with others and it is possible to ship non-pathogenic bacteria (also yeast, worms etc) with specific declarations. You might want to check with your school for a letter and it may be best to have samples shipped to a school address.

Regarding the actual experiment, it sounds like you need the resistance gene on a plasmid which you can then transform competent Agrobacterium with. The transformed bacterium is what will infect the plant and pass on the resistance gene. If you find a lab/mentor that does these kind of experiments, you can also ask them to share a protocol if they send you the bacteria.

Good luck and do reach out again if you have more questions!
MS

Re: Project using Agrobacterium tumefaciens

Posted: Wed Jul 24, 2019 4:56 am
by deleted-738009
Thanks so much for your help!

I found crown gall growing on my rose tree at home so I was wondering if I would be able to cut off the crown gall and isolate a strain of agrobacterium tumefaciens to use??

Thanks so much again!

Re: Project using Agrobacterium tumefaciens

Posted: Wed Jul 24, 2019 9:42 am
by deleted-674917
Hi Kristen,

You are very welcome! Isolating the bacteria yourself for your project is a cool experiment in itself!

I found two links that may be helpful to you. The first is from an undergrad class project and the second is an actual research paper.

https://projects.ncsu.edu/cals/course/p ... rofile.htm

https://scialert.net/fulltextmobile/?doi=crb.2010.27.36

You will need specialized selective medium to grown bacteria from the crown gall isolates so you have to find a way to source that (maybe from a local university lab?)

Hope this is helpful!
MS