IN THIS ISSUE:
Patent awarded for plant gene expression
GMOs under attack in the UK
The farm as factory: Industrial products from engineered crops
Improved regeneration method may speed common bean transformation
A new insecticidal protein to challenge the Bt monopoly?
PTO proposes guidelines for examination of gene and protein claims
Novartis makes major move in agbiotech
Full-text journal articles on the Internet

News and Notes

Delta and Pine Land Company (DPL) and the U.S. Department of Agriculture were awarded a patent on March 3, 1998, for the "Control of Plant Gene Expression" (1). One specific application concerns the control of seed germination. DPL calls this development "Technology Protection" in that the seed of improved varieties, including inbred species, can be produced and/or treated in such a way that the harvested seed will not germinate. The patent work was primarily done by Melvin Oliver, a scientist with the USDA Agricultural Research Service in Lubbock, Texas, through a Cooperative Research and Development Agreement with DPL.

The patented method is based on use of a gene that produces a protein that is toxic or lethal to the plant. One example presented in the patent uses a ribosomal inhibitor protein (RIP) gene. Once produced in a cell, RIP prevents further protein expression in that cell, but is not toxic to other organisms. In the example, expression of RIP is under the control of the LEA promoter, a transiently-active promoter that functions only in late embryogenesis. By placing the RIP gene behind a promoter that restricts its expression to the embryo, only the embryo is affected - it fails to develop.

To allow for normal growth and the production of viable seed, a spacer or blocking sequence is placed between the promoter and the lethal gene in order to keep the gene turned off. On either side of the spacer are placed specific excision sequences that are recognized by a recombinase enzyme whose function is to precisely excise the spacer. The recombinase/excision sequence system can be any that selectively remove DNA in a plant genome. One example given is the CRE/LOX system from bacteriophage. This "second" gene encoding the recombinase enzyme is placed behind a repressible promoter. A third gene encodes a repressor protein which specifically acts on the promoter of the second gene.

Thus, the control of seed germination proceeds by a complex multi-gene, multi-step sequence as follows. A "stimulus" is applied to the plant or seed which blocks the formation of the repressor (turns off the third gene). In the absence of repressor, the recombinase (second) gene is turned on and recombinase enzyme is synthesized; the enzyme chews up the spacer which served to block expression of the RIP gene. With the spacer removed, the lethal gene and its transiently-active promoter are brought back together and RIP is expressed accordingly, i.e., when developing seeds reach the stage of late embryogenesis, and only in the cells of the developing embryo.

Nothing happens without the application of the external stimulus. The stimulus is something to which the plant or plant seed is not normally exposed, such as a chemical (no specific chemical is identified in the patent) or a temperature or osmotic shock. Importantly, this external stimulus does not need to be continuously applied; rather, it can be applied at any time to activate the system. No matter when the system is activated, the plant will grow and produce seed normally. The toxin produced only during late seed development kills the seed embryo; however, the seed remains the same in every other aspect.

In the case of hybrid seed production, a different gene expression strategy is used. One of the parental lines contains the recombinase gene under the control of a promoter that is active right after germination. The transiently active promoter, blocking sequence, and lethal gene are together in the second parent. The hybrid progeny, which are the "technology protected" hybrid seed bought and planted by farmers, thus contain all the elements of the system in every cell. The recombinase, expressed right after germination, proceeds to excise the blocking spacer in front of the lethal gene. Nothing further happens as the seed continues to germinate and grow normally into a mature plant. Not until the plant has flowered and begun to set seed does the last step occur. As before, at the stage of late embryogenesis within the developing seed, the transiently active promoter kicks in and drives expression of the RIP gene and the embryo dies. Thus the seed harvested from the crop will not germinate because it lacks a viable embryo.

There are a number of uses or embodiments described for this patent, and there are 55 claims. These include more secure production of hybrid seed, preventing residual progeny seed from growing as weeds in succeeding crops, preventing the unwanted release of recombinant plant material via seed dispersal, and preventing the saving of proprietary self-pollinated seed.

This last application of the patent's ideas, the production of true-breeding crop varieties, makes use of the "stimulus" system described above. If the external stimulus is a chemical, however, it must be non-toxic to the crop and to animals. An example described in the patent is the Tn10 tet repressor system linked with a modified 35S promoter which is responsive to tetracycline. Tetracycline is an antimicrobial, so its widespread use as a stimulus agent might be controversial.

The genetic system described in the patent has worked to date with cotton and tobacco, but is not completely proven yet. Oliver said that the work has been underway for about five years. The patent was filed on June 7, 1995. He thinks that the tobacco system will be workable within a year, and predicts the cotton system will be ready within two years.

As Oliver points out, control of plant gene expression involves many possibilities besides the control of seed germination. Any plant trait desired in one situation but not in another could conceivably be controlled. Obvious candidates are male sterility, drought or insect resistance, and time of seed germination or flower development.

There are no publications resulting from this work yet due to legal restrictions, according to Oliver; but the patent makes for very interesting reading. It will be several years before Delta and Pine Land Co. will be able to commercialize seed developed with this process, though it is their intention to freely license the technology when they do. Dr. Harry Collins, V.P. of Technology Placement for Delta and Pine Land Co., considers this patent an important tool that will help promote the development of new technologies.

Source
U.S. Patent # 5,723,765; Oliver et al., Control of Plant Gene Expression. March 3, 1998.

Tom Wacek
Urbana Laboratories
twacek@seedsolutions.com

Recent debate about the release of genetically modified (GM) crops into the U.K. environment has been stirred up by none other than the Prince of Wales in an article he wrote for a national newspaper. He claims that scientists are invading territory that should remain the "province of God" alone. Furthermore, the public feels increasingly powerless to demonstrate their concerns about genetic technology. Without adequate labeling of products containing genetically modified ingredients, consumers have no choice and no power to reject the technology.

Consumer groups' apparent victory in the European Parliament, with the introduction of a law which requires the labeling of GM foods, has been short-lived. Loop-holes are now becoming apparent wherein only those foods containing detectable modified DNA or proteins will need labeling. Green and consumer groups are angry, because this will mean that 95-98% of the approximately 30,000 products potentially derived from GM crops will not require labeling.

Green groups have the strong backing of the public. ICM, a market research group, surveyed 500 adults for their views on GM foods. In response to the question: "Do you think crops that have been genetically modified should be kept separate?" 85% said yes, and only 5% said no. "Do you think foods that have been genetically modified should be clearly labeled?" prompted a 96% yes response and only 2% no. In addition, 95% of respondents said yes when asked "Should ingredients derived from GM foods be labeled?" Only 3% said no. A recent poll by another market research organization showed that 70% of British people wanted to ban GM food.

Not surprisingly, industry is concerned at the possibility of a public backlash against genetic engineering technology. Monsanto has launched a new media advertising campaign with the slogan "Food biotechnology is a matter of opinions. Monsanto believes you should hear all of them." The company also has a website to accompany their ad campaign at http://www.monsanto.co.uk/. Each advertisement bears telephone numbers for a number of groups such as Friends of the Earth who are opposed to GM foods. Their position on many of the developments discussed here can be found at http://www.foe.co.uk/camps/foodbio/genepress.html. Some of the green organizations named in the Monsanto ads have complained to the Advertising Standards Agency who deal with unfair advertising in the UK. Green groups claim that they do not have the funds to mount a campaign to counter such huge corporate advertising campaigns.

This frustration has seen some direct-action green groups engage in GM crop vandalism. Early in July, five women were arrested in Oxfordshire for damaging a test site for Monsanto crops. Such protests have recently become more organized. A key project is the "Genetix Snowball" which co-ordinates protests on the first and third Sundays of each month. At each event, a number of protesters turn up and dig up between 1 and 100 GM plants. More and more people are turning up to each event, so while the size of the damage can be considerable, the liability for each individual is minimized -- although the offense of criminal damage can lead to imprisonment.

A spokesperson for Monsanto has said that such activities are "just plain vandalism." There is also the concern that any safety protocols put in place by the crop growers will become meaningless if plants are carried off-site. Activist Zoe Elford maintains that all transgenic plants are bagged up and left on site. Currently, the locations of all GM crop test sites are available from the government. This makes life for the protesters somewhat easier. However, Monsanto argues that making such information confidential will not stop the determined few who seek to damage GM crops. Monsanto insists that the most appropriate way of challenging GM technology is by engaging in a debate with the government, not by attacking the companies or research establishments which are doing the actual field testing.

However, any challenges to current government policy have been dealt a severe blow this month with the case of Guy Watson, an organic farmer from Devon in the southwest of England. Supported by the Friends of the Earth and the Soil Association, Watson failed to convince a court that it was necessary to seek a judicial review of GMO trials in the UK. He is concerned that cross-pollination of his organic sweet corn crop by GM maize would lose him his organic status and be financially disastrous for his business. The judge, saying Watson's case was "unarguable," ruled that the government was entitled to accept expert advice that the risk of genetic contamination from the transgenic maize strain was "likely to be zero." However, it has been reported that representatives of industry have stated that "those who want guarantees of totally GM-free food must compromise and accept some contamination."

Contamination of any degree is not acceptable to organic food growers who must meet European standards in order to sell their produce as "organic." These standards include the requirement that there is no GM material in the plants. The Soil Association is one of the organizations which grant organic status in the UK. They argue that it will become impossible to consider any crops as organic. The Guardian newspaper reports that Watson has said, "If the trials are successful and the seeds get on to the national seed list, genetically modified sweet corn will be grown throughout the south of England. Every July and August, the air will be saturated with GM pollen. It will be impossible to grow an organic crop."

The journal Nature (July 2, 1998) reports that the actual effects of cross-pollination are difficult to assess, because the probability of such events is so small. While Pete Riley of Friends of the Earth states that pollen can maintain its fertilizing ability for up to 80 hours after flowering, government experts say that the "vast majority" of pollen grains can only fertilize for up to half an hour. Furthermore, it has been internationally agreed that 99.9% seed purity can be achieved by maintaining a 200 meter buffer zone between crops. In the case of Mr. Watson, the distance between the organic and the GM crops is 2 kilometers. Other factors which make cross-pollination exceedingly unlikely include the low probability that the two corn crops will flower at the same time, and the need for the organic crop to be downwind of the GM corn in order to be pollinated by it.

In response to the case, the Supply Chain on Modified Agricultural Crops (Scimac) has suggested that dialogues be opened between organic farmers and growers of GM crops. Where field arrangements cannot be reached (bearing in mind that GM rapeseed can transfer pollinating material over 4 km), more agreeable solutions might be achieved by the use of "mule" crops that cannot propagate, and "terminator" technology which involves engineering crops so that their seeds will not germinate. There is, however, some resistance to this latter option, because it would require farmers to purchase new seed each year.

By now the sweet corn crop will have reached the stage of pollination. The UK government will have to respond promptly if confidence is to be maintained in the organic food industry.

Sources
This article was drawn from articles published in London and regional editions of The Daily Mail, The Guardian, and The Observer, during the period June 6 - July 11, 1998.

Ben Thomson
Department of Law
University of Sheffield
lwp97bt@sheffield.ac.uk

Plant Research News

Agriculture's traditional role in providing food, feed, and fiber is being expanded by biotechnology into entirely new forms of production. Some farms in the future will be living factories churning out industrial chemicals from genetically engineered crops. Research projects involving scientists from Sweden, Australia, and England have developed plants producing unusual oils used in the production of polymers, plasticizers, lubricants, and other industrial products, thus providing a renewable alternative to petrochemical oil.

Acetylenic and epoxy fatty acids are critical raw materials used in the production of polymers such as plastics and certain chemicals. These fatty acids, which are modified forms of those present in edible oils, are currently derived from either non-renewable petroleum or chemically processed vegetable oils. Many wild plants, such as various species of the genus Crepis, a member of the Compositae or sunflower family, are known to produce these unusual fatty acids. A paper in the journal Science describes the cloning and expression in transgenic plants of genes involved in the synthesis of acetylenic and epoxy fatty acids (1). The chemical modifications of oil are done within the plant, obviating the need for expensive industrial processing and eliminating waste.

An epoxygenase gene cloned from C. palaestina encodes an enzyme responsible for making epoxy fatty acids, which are major components in the production of polymers used in manufacturing adhesives. The acetylenase gene, cloned from C. alpina by the Swedish team, produces acetylenic fatty acids which have potential applications in the synthesis of high quality surface coatings and ester-type lubricants. The genes were isolated from the plants' seed cDNA libraries using PCR primers specific to desaturase genes. Both epoxygenase and acetylenase enzymes appear to be non-heme diiron proteins with histidine rich motifs, according to the report. "This is the first time in the world anyone has isolated a gene responsible for the production of an acetylenic compound. Nobody even knew that such a gene existed before our work started," says Sten Stymne of the Swedish University of Agricultural Sciences.

Seeds from transgenic Arabidopsis plants expressing these genes contained up to 15% by weight epoxidated vernolic fatty acid or up to 25% acetylenic (crepenynic) acid. The control plants did not contain even a trace of these fatty acids. Allan Green of the Commonwealth Scientific and Industrial Research Organisation (CSIRO) says that "the possibilities are immense -- components of detergents, nylon, glue, paints, lubricants, and plastics could all be produced from plants, rather than fossil materials. Plants could provide a renewable, biodegradable source of these high-value specialty products."

While the research shows that industrial feedstocks can be produced in plants, Green says that the plants must produce much higher levels of these fatty acids to be commercially viable. The group now is trying to identify additional genes responsible for the high accumulation of epoxy and acetylenic acids in wild plants. They are also cloning genes from plants producing oils which have epoxy groups in the various positions in the fatty acid chain and identifying active sites of these enzymes.

This work may help in the production of diverse chemical feedstocks, according to Surinder Singh of CSIRO. Although the initial transgenic research was done in Arabidopsis, the researchers believe that specialty oils eventually will be produced in flax (linseed), an industrial crop whose oil has the ideal starting composition. To minimize the risk of contaminating edible oils through gene flow, Green says that it is better to target self-pollinating oil crops such as flax rather than outcrossing ones such as canola or sunflower. He anticipates that it may be another five to eight years before crop "mini-factories" are producing high value industrial compounds down on our farms.

Source
Lee, M., M. Lenman, A. Banas, M. Bafor, S. Singh, M. Schweizer, R. Nilsson, C. Liljenberg, A. Dahlqvist, P. Gummeson, S. Sjodahl, A. Green, and S. Stymne. 1998. Identification of non-heme diiron proteins that catalyze triple bond epoxy group formation. Science 280:915-918.

C. S. Prakash
Center for Plant Biotechnology Research
Tuskegee University
prakash@tusk.edu

The common bean (Phaseolus vulgaris) is the most important food legume for more than 300 million people, most of whom live in the developing world. According to the International Center for Tropical Agriculture (CIAT) in Cali, Colombia, it is the second most important source of protein in eastern and southern Africa and fourth in tropical America, where the crop was first domesticated.

One of the vexing characteristics of raising this important crop has been the perceived necessity of spraying fungicides and insecticides "regardless of the insect situation," in high amounts and with great frequency, in some cases reaching up to 24 applications in a crop cycle of 90-100 days in Colombia. In Peru, where pesticides are costly, insect control may reach 23% of the farmer's total costs (1). These practices, which challenge the bean farmer's budget, present health problems in farming communities as well.

Given the above circumstances, developing pest-resistant transgenic common bean varieties is of high importance. But the efficiency of genetic transformation protocols depends on in vitro regeneration methods, and those used for other plants do not produce desired results with the common bean. At the International Symposium on Breeding of Protein and Oil Crops held April 1-4, 1998 in Pontevedra, Spain, a joint team including A. Mejía Jiménez and co-workers from both CIAT and Germany's Hanover University, reported on a regeneration system for the common bean that may prove useful for genetic transformation (2).

Using genotypes of agronomic importance, the team was able to induce a highly regenerable tissue that can be maintained undifferentiated indefinitely. The latter characteristic is important as it "allows more efficient in vitro selection," said Mejía Jiménez in a recent interview. He described this tissue as a "non-traditional callus, with minimal differentiation and high organization during its growth." They named the tissue meristematic callus, or m-callus, as it is composed solely of nude meristems lacking in differentiated organs like primordial leaves, buds, or shoots.

These meristems are not formed de novo, but arise from those already present in the cotyledonary node of the initial explant. They are induced by Thidiazuron (TDZ), a growth regulator with strong cytokinin-like activity. The m-calli can then reproduce in vitro indefinitely, and the "weaker" cytokinin 2iP is used to bring on shoot differentiation. The shoots, which tend to be difficult to lengthen, root, and establish outside of tissue culture conditions, are then micrografted onto hypocotyl segments of seedlings germinated in vitro. This procedure results in elongated and rooted specimens that can be transferred to the greenhouse within six weeks.

Mejía Jiménez points out that a transgenic variety of the common bean has yet to appear on the market. He and his colleagues hope the m-calli-based regeneration system they have developed will quicken the arrival of bean transformation through Agrobacterium or particle bombardment, beginning with "varieties resistant to pathogens, which would allow for a significant reduction in the use of agrochemicals."

Sources
1. Annual Report, CIAT Bean Program, 1994 (For Internal Circulation).

2. Mejía Jiménez, A. et al. 1998. Development of an in vitro regeneration system in common bean suitable for genetic transformation. In: Antonio M. de Ron, Coordinator, Proceedings, International Symposium on Breeding of Protein and Oil Crops, Pontevedra, Spain.

Timothy Pratt
Journalist
Cali, Colombia
thipra@norma.net

The most notable success of agricultural biotechnology so far has been the development of insect-resistant crops, which were planted on 10 million acres in 1997. These crops garnered $300 million in benefits from increased productivity and cost-savings due to reduced pesticide use. The pest-fighting ability of these crops is powered by a handful of insecticidal genes isolated from a single bacterium, Bacillus thuringiensis. This precarious dependence on a sole source of pest resistance has led to fears that insects will eventually adapt to Bt genes. There is clear experimental evidence to validate such a concern (See July 1997 ISB News Report).

A new insecticidal toxin has been discovered that may challenge the Bt monopoly. A team of scientists from the University of Wisconsin-Madison, led by Richard ffrench-Constant and David Bowen, has discovered new toxins from a bacterium that may represent the "next generation of microbial insecticide" (1, 2). The gram-negative bacterium Photorhabdus luminescens packs a considerable arsenal within its cell: toxins, antibiotics, antifungal compounds, lipases, proteases, and even light-producing genes. The bacteria thrive inside the gut of an insect-attacking nematode. When the nematode invades an insect host, it releases the bacteria into the insect's hemocoel. The bacteria then kill the insect, leaving a cadaver that eerily glows in the dark! The nematodes then eat both the bacteria and insect carcass, with hundreds of nematodes eventually bursting out of a single insect victim. "This makes Alien look like a cakewalk," says ffrench-Constant.

Photorhabdus "is a voracious pathogen. One bacterial cell can kill an insect," says Jerald Ensign of UW-Madison, who with then-graduate student Bowen, discovered the toxic potential of this bacterium which kills its host in 24 to 48 hours. Even picomolar quantities of Photorhabdus toxin can be lethal to many pests such as caterpillars, mealworms, and even cockroaches and ants.

The UW team isolated a toxic protein fraction from Photorhabdus and found that it had four protein components. When fed to tomato hornworm, complexes A and D showed very high toxicity. Bowen, Michael Blackburn, and Thomas Rocheleau in the ffrench-Constant lab then probed an E. coli expression library of Photorhabdus genes with antisera raised against the toxin, and cloned the genes encoding the four proteins. Disappointingly, toxin proteins expressed in the E. coli system were not secreted and were not toxic. Deletion mutants of Photorhabdus, however, confirmed that tca and tcd genes encode orally active toxins. The mode of action of these toxins is not yet known and it also remains to be established that transgenic plants expressing these genes indeed become resistant to insects.

The response of the scientific community to the new toxin has been positive but cautious. Fred Gould of North Carolina State University, who studies the evolution of insect resistance to Bt, welcomes the new report. "Finding a totally new toxic protein is good news from the perspective of resistance management. If the two genes can be expressed together in engineered plants, this could be used advantageously." As these toxin proteins would be expressed inside the plant, there should be much less exposure of parasites and predators to them than there has been to conventionally sprayed insecticides, he adds.

Bruce Tabashnik of the University of Arizona, also a Bt-resistance researcher, says that the new discovery is very exciting but notes that "much work will be needed to determine the practical value of these new toxins. Knowledge of their spectrum of toxicity against insects and other organisms will be essential. Also, it remains to be seen if these toxins are insecticidal when expressed in plants, particularly in light of their lack of toxicity when expressed in E. coli. In the meantime, we must use Bt wisely -- both to prolong its efficacy and to establish methods for delaying pest resistance that can be applied to new toxins."

Rick Roush of the University of Adelaide in Australia says, "We definitely need alternative toxins to Bt and the Photorhabdus toxins may be good candidates. However, there is also a need to temper some of the enthusiasm because Photorhabdus toxins are much more poorly known than Bt toxins and because they have broader activities, safety testing must be very extensive before they are used in transgenic crops." He further adds, "It is important to remember that in spite of how well Bt toxins had been characterized by the mid-1980s, it was more than ten years from the production of the first Bt transgenic plant to the commercial introduction of Bt crops in 1996. Photorhabdus toxins may eventually help us to manage resistance to Bt toxins, but for at least the next ten years, we need to be planning to manage Bt resistance without them." Neal Stewart of the University of North Carolina at Greensboro concurs, and says that the broad-spectrum activity of Photorhabdus "could have numerous ecological effects from food web interactions to decreasing beneficial insect populations. Its effects will need to be tested empirically."

In response, ffrench-Constant agrees that further work is necessary but points out that Photorhabdus "is one of the only viable alternatives to Bt out there." The UW group is trying to boost the secretion of the Pht toxin and also testing its safety on humans and wildlife (2). The Photorhabdus technology has been licensed to the Indianapolis-based Dow AgroSciences.

Sources
1. Bowen, D., T. Rocheleau, M. Blackburn, O. Andreev, E. Golubeva, R. Bhartia, and R. ffrench-Constant. 1998. Insecticidal toxins from the bacterium Photorhabdus luminescens. Science 280:2129-2132.

2. Strauss, E. 1998. Possible new weapon for insect control. Science 280:2050.

C. S. Prakash
Center for Plant Biotechnology Research
Tuskegee University
prakash@tusk.edu

Microbial Research News

Under U.S. patent law, a patent application must contain a written description of the claimed invention. One objective of this requirement is to assure that the inventor makes it clear that he or she possessed the claimed invention when the application was filed. But just how much information must a patent application contain to show that an inventor possessed a claimed gene or protein?

In 1993, the Court of Appeals for the Federal Circuit decided that a patent application did not contain a written description of a human beta-interferon gene because the application did not provide its nucleotide sequence (1). Although the patent application taught that a disclosed cDNA clone could be used to isolate human beta-interferon mRNA, the court asserted that a description of the DNA itself was required, not just a potential method for obtaining the DNA molecule. Last year, the Federal Circuit reiterated its position that a description of a method for obtaining a gene does not provide a written description of the gene itself (2).

In a case before the U.S. Patent and Trademark Office (PTO) Board of Patent Appeals and Interferences, Andrew Baird had described more than a method for isolating a bovine fibroblast growth factor gene; his application had disclosed the bovine FGF amino acid sequence and a nucleotide sequence that would encode the FGF (3). Yet the Board concluded that Baird had not met the written description requirement for the naturally occurring bovine FGF gene, because his patent application did not teach the particular nucleotide sequence of the naturally-occurring gene.

Taking it one step further, suppose that you have a patent application that provides the nucleotide sequence of a new soybean storage protein gene. This information would support a claim to the new gene itself. But suppose that you wish to expand patent protection to cover synthetic variants of the storage protein gene, as well as homologues that you predict reside in faba beans, peas, and broadbeans. How much additional information will you need to obtain a genus claim to the storage protein gene?

In the Baird case, the Board decided that a description of bovine FGF does not provide a written description for the genus of mammalian FGFs. More recently, the Federal Circuit decided that a disclosure of the nucleotide sequence for rat insulin did not support a claim to the genus of vertebrate insulin genes or mammalian insulin genes (2). The decision does not make it clear, however, just how many genes one must disclose to describe a genus of related genes.

The PTO tackles questions about biotechnology-related species and genus claims in its proposed guidelines for determining compliance with the written description requirement. According to the guidelines, there are two basic ways to describe a particular gene or protein. The first way is to disclose its complete structure. For example, the disclosure of the complete nucleotide sequence of a gene would support a claim to the gene, as indicated by case law.

In the absence of a complete sequence, a patent application must provide other identifying information, such as physical characteristics, or functional characteristics that can be coupled with a correlation between function and structure. As an illustration, the guidelines state that a DNA molecule would be sufficiently described by including information about its length, origin, and restriction endonuclease sites. In another example, the PTO deems a claim to an isolated alginate lyase enzyme to be supported by a description of the enzyme's molecular weight, origin, activity, and specificity.

With regard to genus claims, the guidelines indicate that a patent application must provide a description of a "representative number" of adequately described species. The number of species that must be disclosed depends upon the amount of variation within the genus. Therefore, it may be necessary to provide a written description for each member of the genus if the members are expected to vary considerably in their identifying characteristics. Note that the PTO considers a general description of a genus, such as "vertebrate insulin cDNA," to fail the written description requirement, because the members of the genus are defined only by function.

On the positive side, the proposed guidelines do indicate that one approach to obtaining a claim to a genus of DNA molecules is to define its members by hybridization properties. For example, a patent application could disclose the nucleotide sequence of a reference DNA molecule and provide several examples of other DNA molecules that hybridize with the reference DNA molecule under specific conditions. At this time, the PTO seems to require that such a genus claim must include the details of both hybridization and washing conditions.

Although not mentioned in the proposed guidelines, PTO examiners are also considering nucleic acid molecule and polypeptide genus claims in which members are defined in terms of sequence identity to a reference molecule. For example, one could claim polypeptides that have an 85% amino acid sequence identity with a disclosed storage protein amino acid sequence. Currently, the PTO seems to have a policy that such a claim must describe the algorithm and the parameters that one would use to determine sequence identity.

The PTO is accepting comments from the public on the proposed written description guidelines until September 14, 1998. Hopefully, these efforts will provide an approach for examination that is consistent with the state of the art, and that examiners can implement in a predictable manner.

Sources
1. Fiers v. Sugano, 25 USPQ2d 1061 (Fed. Cir. 1993).

2. The Regents of the University of California v. Eli Lilly and Company, 43 USPQ2d 1398 (Fed. Cir. 1997).

3. Fiddes v. Baird, 30 USPQ2d 1481 (BPAI 1993).

4. The proposed guidelines for biotechnology-related species and genus claims were published in the June 15, 1998 Federal Register; they are also available on the PTO home page
( http://www.uspto.gov/web/offices/com/sol/notices/fr986-27.html).

Phillip B.C. Jones, Ph.D., J.D.
Seattle, Washington
pbcj@emeraldnet.net

Industry News

Not to be left out of the recent flurry of agbiotech corporate activity, Novartis announced the planned investment of $600 million over the next ten years to fund one of the largest initiatives in plant genomics. The first step will be the creation of the Novartis Agricultural Discovery Institute (NADI), which will be one of the largest single research endeavors dedicated to agricultural genomics research and development. Located in San Diego, California, the main campus of NADI will have a team of about 180 researchers in 50 laboratories.

NADI researchers will apply genomics technologies to the development of improved plant traits, new methods for crop protection, and new animal health products. NADI will focus on matching genes with traits, primarily through the production and exploitation of gene and protein databases, supported by tools for protein function and structure analyses, and engineering. Initially, NADI will explore plants, fungi, bacteria, viruses, nematodes, and insects, including some model systems

NADI will be a cornerstone of Novartis' biotechnology research, designed to allow cooperation between other Novartis groups including Crop Protection and Seeds. It will work in tandem with the Novartis Agribusiness Biotech Research facility at Research Triangle Park, North Carolina, and with numerous Novartis research stations worldwide. The placement of NADI near the recently announced Novartis pharmaceuticals genomics institute (Novartis Institute for Functional Genomics), which is being built in La Jolla, has the goal of optimizing cross-business synergies in genomics research in both agribusiness and in pharmaceuticals.

The planned investment begins with an initial phase involving $250 million, to be used in part for the establishment and operation of the NADI, a wholly-owned entity of the Novartis Research Foundation. Approximately $50 million will go towards the building of NADI, with another $55 million anticipated for annual operating budget. Building is scheduled to begin in 1998, with completion anticipated in 1999.

Sources
1. Novartis announces $600 million investment in agricultural genomics. Novartis home page (http://www.novartis.com), July 1998.

2. Welch, M., Novartis Earmarks $600 million for agricultural genomics. BioWorld Today, Vol. 9, No. 140, July 23, 1998, pp. 1,6.

William O. Bullock
Institute for Biotechnology Information, LLC
Research Triangle Park, NC
http://www.biotechinfo.com

Internet News

The information superhighway has done much lately to divert traffic away from the campus library. Many scientists and students visit the virtual library right on their desktops to read scientific papers in their favorite journals. Whether you are in Topeka or Tbilisi, browsing through a new scientific paper in the magazine Science is just a few mouse clicks away! Most publishers are racing to place their journals on the Internet where one can not only read complete scientific articles but also jump to cited papers in the title, connect to the nucleic acid or protein databases, and even e-mail the authors instantly. The awesome power of the Internet thus goes beyond providing simple electronic access of published text to pack more punch in journal reading.

Here I list a few Internet sites to retrieve full texts of scientific articles in biotechnology from the peer-reviewed print journals. Almost all sites require a fee for such access although most let you browse their table of contents (TOC), abstracts and some general articles for free. A few journals have free trial periods while many publishers provide free "TOC alert" whereby the titles from your favorite journals are emailed to you periodically.

A comprehensive listing of many molecular biology journals online can be found at the "Cell and Molecular biology Online" site ( http://www.cellbio.com/elecpubs.html) and at "Biochemistry and Molecular Biology Journals" (http://www.geocities.com/~jrbeasley/biochem/journals.html).

The most innovative and thoughtful effort in placing scientific journals on the Internet is from Stanford University, called HighWire Press (http://highwire.stanford.edu/). To reduce exploding journal costs and to make use of the communication opportunity provided by the Internet, HighWire has teamed up with many non-profit journal publishers such as scientific societies and universities to provide electronic access to journals such as:

Science (http://www.sciencemag.org/)

PNAS (http://www.pnas.org/)

Cell (http://www.cell.com/)

Genetics (http://www.genetics.org/)

The EMBO Journal (http://www.emboj.org/)

The Plant Cell (http://www.plantcell.org/)

Plant Physiology (http://www.plantphysiol.org/)

The latter two are available without charge through 1998. While at the HighWire site, be sure to visit their section on "Tips for Better Browsing" (http://highwire.stanford.edu/tips/) for some very useful advice such as increasing the font size and viewable area on your browser and how to stop those distracting spinning graphics.

The commercial publishing houses also provide electronic access to their journals but the price of such access can be steep. However, if your library has a subscription to the print version of a particular journal, you may be able to access the electronic version at no cost. Elsevier, which publishes Biofutur, Biotechnology Advances, Trends in Biotechnology, and other journals of interest to biotechnology has perhaps the best site with access to nearly 1000 journals (http://www.sciencedirect.com). The Springer site (http://link.springer.de/) lets you read journals such as Plant Cell Reports, Theoretical and Applied Genetics, Crop Science, Applied Microbiology & Biotechnology, and Molecular & General Genetics. The Academic Press journals are available at http://www.idealibrary.com/ while the Kluwer site (http://www.wkap.nl) has Plant Molecular Biology, Plant Molecular Biology Reporter, and Molecular Breeding although their Transgenic Research and Euphytica are not yet on the Internet.

Most journals now offer their papers in both HTML and PDF versions. The PDF version lets you print the journal article in exactly the same format as the original print version and requires that your computer has the Adobe Acrobat software, which can be downloaded for free at http://www.adobe.com/prodindex/acrobat/readstep.html. By using a color inkjet printer and glossy paper, one can print scientific papers identical to the original reprints right in your office complete with graphics and half-tone photographs. The HTML format is intended for desktop viewing and offers additional features such as hyperlinks within a title to tables and figures (which could be zoomed up), and to other cited and related papers through the Medline database.

Some very high-impact biotechnology journals from the Nature group (http://www.nature.com/) including Nature Biotechnology (http://biotech.nature.com/) offer TOC, abstracts, and news articles on their sites, but do not yet provide full texts of journal articles on the Internet.

Biotechnology is a highly information-intensive science and the availability of scholarly journals now on the Internet has democratized access to scientific information. Hopefully this access will become more affordable for individual academic users as the number of such users increases over time.

C. S. Prakash
Center for Plant Biotechnology Research
Tuskegee University
prakash@tusk.edu

The material in this News Report is compiled by NBIAP's Information Systems for Biotechnology, a joint project of USDA/CSREES and the Virginia Polytechnic Institute and State University. It does not necessarily reflect the views of the U.S. Department of Agriculture or of Virginia Tech. The News Report may be freely photocopied or otherwise distributed without charge. P.L. Traynor, Editor.

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