Frequently Asked Questions

See Also Frequently Asked Questions about Request Procedures or download the TransBacter Request Form.

What is TransBacter?

TransBacter is a collective name given to non-pathogenic bacteria that have been modified to be able to replace Agrobacterium in plant transformation processes. To date we have shown, using modified plasmids, that three species of bacteria, Sinorhizobium meliloti, Mesorhizobium loti, and Rhizobium sp. NGR234 are able to transfer genes to plants under certain circumstances.

TransBacter strains have been made available to the scientific community through a BiOS License.

By using TransBacter it may be possible to circumvent the current licensing thicket involved with using Agrobacterium for plant transformation (see Agrobacterium-mediated Transformation of Plants Technology Landscape for more information).

How can I obtain TransBacter strains?

Unfortunately, we no longer provide TransBacter strains.   However, as Cambia is placing its technology in the public domain, you may now request any of our vectors from labs that use or published using them.  Cambia is Ok with this arrangement and does not require any payment or license.  While we will not assert our rights over any of the pCAMBIA vectors or technologies, there may be other rights held by third parties associated with components or particular uses.

You may also need to find out about requirements for importing the bacteria into your country. CAMBIA doesn’t accept any liability on the vectors or for checking these because each jurisdiction and institution may have different rules, and complying with these is the responsibility of the recipient. However, if you need information to fill out forms, you may be able to find it on this website (see below) or ask colleagues from your country through our discussion forum. We encourage people who have imported these bacteria to post the information they used.

Is TransBacter patented or owned by a company?

The patent applications and trademark owned by CAMBIA are being placed in the public domain. CAMBIA did not receive any profit from selling, using, or out-licensing TransBacter. The technology was patented at CAMBIA’s expense in order to manage the rights to practice the technology in accordance with the principles of “open source”. An entity may obtain a license to use the technology only by agreeing to all the terms of the license, including allowing other licensees to use improvements and biosafety data. An entity that attempts to enforce patents on the improvements against others voids its license and would be liable for infringement.

What are the BiOS License conditions relevant to TransBacter?

In summary, in return for CAMBIA’s permission to use the technologies, a licensee institution was to allow and encourage its employees and students to post any improvements made to the technologies on this website. The licensee institution also agrees not to assert any intellectual property rights to the improvements against other licensees.

All licensees that comply with these license terms were allowed to use the technology for research, public good, and/or commercial product development.

Companies with for-profit governance also were expected to assist us in meeting costs such as providing this website and any improved protocols that are developed. However, CAMBIA’s BiOS license does not require any commitment to repay patenting costs, nor any milestone payments or royalties.

How much does it cost?

All BiOS licenses were completely royalty-free.

How can I obtain a license to use TransBacter strains in a for-profit or commercial environment?

There is no longer a requirement to have a license to use TransBacter strains for both for-profit business or non-profit research.

Old copies of both the BiOS License for Plant Enabling Technologies and the attached Technology Support Agreement are available at BiOS website.

Will I have to pay royalties to use TransBacter?

BiOS licenses are available from CAMBIA completely royalty-free for both research and commercial purposes.

Does CAMBIA make a profit from the licenses?

Very few companies make a profit on patent licenses! Patents are expensive to file and maintain. CAMBIA did not make a profit with its licenses. We are a non-profit organisation.

How do I register as a non-commercial BiOS Licensee of TransBacter on BioForge?

No need to register.

Can Transbacter be used to make a profit?

Royalty-free licenses are available from CAMBIA to both for-profit and non-profit entities. CAMBIA believes that hunger in many parts of the world can be alleviated not only by providing tools to public good researchers in those communities, but also by removing barriers to micro, small and medium enterprise.

IBM, Nokia, Sun and other IT companies have shown that hardware and software technology can be non-exclusively and open-source licensed and simultaneously generate profitability in a for-profit company. BiOS-compliant licenses make it possible to do the same for life sciences products.

My friend works in a lab that has TransBacter strains. Can I ask for the strains? Do I still need to pay?

Yes, you can request to use strains from your colleague and Cambia does not require any payment or license anymore.

How do I import bacterial strains into my country?

Every jurisdiction has different rules. You will need to find out if there are any importation/custom requirements for mailing of these strains to your country.

Once you have obtained the appropriate forms, these frequently asked questions and other information on the website should give you enough information to fill out the forms. We encourage people who have imported these bacteria to post the information they used.

For example, see General Information about TransBacter which contains the information that was required by a recipient within the US to successfully obtain a USDA-APHIS permit.

What are the exact constituents of the kit and their quantity?

Depending on your request from a lab, the kit will contain bacterial strains (e.g. Sinorhizobium meliloti, Rhizobium sp. or Mesorhizobium loti). Each bacterial strain can be individually streaked onto 1.5mL of YM Agar medium in a sealed plastic tube.

The number of tubes containing bacteria on media depends on the number of bacterial strains requested.

What binary vectors are available to use with TransBacter?

pCAMBIA1105.1R is specifically provided to use with TransBacter strains. The “R” signifies the presence of a multiple cloning site with a size marker that distinguishes it from pCAMBIA1105.1. Some variants of this vector are now available. pCAMBIA1105.1 is intended to use with Agrobacterium controls. A description and maps of these vectors can be downloaded here.

What introduced genetic constituents do the strains contain?

The strains we are distributing are modified in that they contain a derivative of the pTiBo542 plasmid from Agrobacterium tumefaciens. pTiBo542 had been “disarmed” by having both of its T-DNA regions deleted, to create pTiEHA101 (with an nptI gene in place of the deleted T-DNA) and pTiEHA105 (with no foreign DNA inserted in place of the deleted T-DNA). For details, see Hood et al 1986 J Bact 168:1291-1301 and Hood et al 1993 Trans Res 2:208-218.

pTiEHA105 was modified at CAMBIA by homologous recombination insertion of a pCR2.1TOPO plasmid (Invitrogen Inc.) containing an RK2 oriT (amplified from pSUP202 with primers EVS49 and EVS50 – see Stabb and Ruby 2002 Methods Enzymol. 358:413-426) and the virG gene – to create pTiWB1 – or the moaA gene – to create pTiWB3.

Some strains also contain the binary vector pCAMBIA1105.1R with a GUSPlus reporter construct (see the GUSPlus Project).

Can the strains be obtained without a binary vector?

Several strains and combinations are now available from various labs around the world.

Why can’t I get Mesorhizobium loti or Rhizobium sp. NGR234 without pCAMBIA1105.1R in it?

In both cases the strain was first transformed with pCAMBIA1105.1R by electroporation and the modified Ti plasmids pWBTi1 or pWBTi3 were introduced next by triparental mating from Sinorhizobium meliloti.

You should be able to cure these bacteria of the binary plasmid by growth in non-selective conditions (no spectinomycin or streptomycin) for some generations, followed by replica plating on media with and without spec (or strep) to screen for colonies that have become sensitive to it.

A quicker method would be to transform in another pCAMBIA plasmid (or other plasmid with the same backbone such as the pPZP plasmids) and select for its resistance gene. This should “bounce out” pCAMBIA1105.1R since cells don’t like to keep two different plasmids with the same replicon. You should still screen for loss of spec/strep resistance though, to make sure.

Can I use TransBacter to transform my species of interest?

We have experience using TransBacter to transform tobacco, Arabidopsis and rice. Other labs have had similar success and are trying other species. We encourage you to use TransBacter on the organism that you are researching.

I use TransBacter in my lab. Can I help distribute it?

Your state or country may require permits for the strains. The conditions of the BiOS license require that recipients comply with all applicable laws.

We try whenever possible to refer requestors to a BiOS Reference Lab in the same nation for the redistribution of materials. Depending on the applicable laws in your nation, this may help to minimise import and quarantine restrictions. If you have the strains or vectors and would like to be a BiOS Reference Lab in your country to replicate and send out the strains on our behalf, please e-mail us.

How else can I help?

• Become a BiOS Reference Lab in your country and help us distribute strains.
• Use TransBacter
• Tell your colleagues about TransBacter and encourage them to use it in their research.

For more information about how you can help, use the links on the left under “How do I contribute?”