New Eukaryotic GUS Gene “FunGus”

A fungal beta-glucuronidase has been discovered, described in the patent publications US 2005/153448 and WO 2005/68617 A2, and in a recent publication. Patent claims have recently issued in the United States in US Patent 7,148,407. This gene has closer codon usage to that of other eukaryotes than that of bacterial glucuronidase genes.

We have successfully used this gene in reporter constructs, pCAMBIA0306.2 and pCAMBIA1306.2 (see VectorNTI diagrams attached–download using VNTI Viewer), for screening transformation in rice.

Patents are about to issue on this gene. Accordingly, these constructs will become available from CAMBIA in early 2008 under the same terms as the GUSPlus constructs described on other BioForge pages, i.e. under the BiOS License.

See page describing the nomenclature of pCAMBIA vectors available from CAMBIA.

Why should I use FunGUS?

The FunGUS protein is expressed naturally in a eucaryote, unlike Escherichia coli GUSA. Like GUSPlus, it also has the great advantage in that it can be secreted by plant cells (the gene is fused to a sequence encoding the glycine rich protein secretion signal peptide which results in the GUSPlus protein being efficiently transported to the periplasmic or apoplastic space, where it is active). This allows non-destructive in vivo staining and detection in living tissues that can be grown further after staining. Also, FunGUS is being offered widely to both non-profit and for-profit organizations that are willing to agree to the terms of a BiOS license. Under this license, the genes may be used in research or for commercial product development royalty-free and without payment of additional license fees, but only under the condition that all improvements are granted back to the common group of licensees. This license structure, pioneered within the BiOS Initiative, is to ensure that a choice by any licensed innovator to use these genes has the best chance to lead to deliverables, as opposed to becoming encumbered in patent thickets that prevent productive use.

How can I obtain pCAMBIA vectors containing FunGUS?

pCAMBIA FunGUS Vectors (pCAMBIA0306.2 and 1306.2) are currently unavailable but we aim to provide as soon as we have the resources available. These plasmids will be available under CAMBIA’s BiOS license or a BiOS-compliant MTA. Contact us at materials@cambia.org.

Will I have full freedom to use pCAMBIA vectors?

Without knowledge of the detail of what you are trying to do it is not possible to make any assertions about this, and CAMBIA cannot warrant it. Many aspects of what you may want to do, for example the use of Agrobacterium to transform plants, and some DNA sequences within the pCAMBIA vectors, may be covered by patents issued to other entities in many countries. Although you may have a research exemption (either de facto or de jure) or certain aspects of your work may not be covered by a patent in your country, it could be subject to patents where products developed from your work might later be used or exported. In such circumstances the owner(s) of the patent rights might legally prevent deliverability of the results of your research work. This is one of the most important reasons for the BiOS license developed by CAMBIA, and available under the same license CAMBIA has developed and is improving a plant transformation system that will be less limited by restrictive licensing of patented technologies. If you want to learn more about this project, or join in the community of people trying to make it happen, please visit the TransBacter project. pCAMBIA0306.2 lacks the hygromycin resistance gene covered by Syngenta patents in some jurisdictions, and was designed for use in screening for transformants non-destructively without antibiotic selection. We have successfully used it in this way for obtaining transformants in both high-frequency and low-frequency plant transformation systems, notably for rice transformation.

What are the BiOS License conditions relevant to FunGUS?

In summary, in return for CAMBIA’s permission to use the technologies, a licensee institution agrees 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 are allowed to use the technology for research, public good, and/or commercial product development. Companies with for-profit governance also agree 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.

Will I have to pay royalties to use FunGUS?

BiOS licenses are available from CAMBIA completely royalty-free for research and commercial purposes. For-profit companies based in OECD countries such as Australia and the USA are normally requested to enter into a technology subscription agreement, or a similar arrangement that helps CAMBIA defray costs of keeping the technology available on the BioForge. For-profit companies based outside the OECD can take up such a subscription for a reduced fee or in-kind assistance (please contact us for information), and non-profits obtain all the benefits of the license and technology support subscription without paying a fee. CAMBIA has committed to this broadly accessible licensing arrangement in the hope that many who wish to see the protected commons of improvements expanded for public good will use this technology. The patents covering FunGUS are owned by CAMBIA, which is a non-profit registered in Australia and a non-government organisation registered with the Food and Agriculture Organization. CAMBIA does not receive any profit from selling, using, or out-licensing FunGUS. The technology has been patented at CAMBIA’s expense in order to manage the rights to practice the technology in accordance with the principles of “open source”. Any entity may obtain a license to use the technology only by agreeing to grant back improvements to all other licensees. The same license allows the use of CAMBIA’s GUSPlus, transactivation and Transbacter technologies (see other projects on this BioForge website).