‘It has been shown that glyphosate tolerant plants can be produced by inserting into the genome of the plant the capacity to produce a higher level of EPSP synthase in the chloroplast of the cell…which enzyme is preferably glyphosate tolerant. Variants of the wild-type EPSPS enzyme have been isolated which are glyphosate tolerant as a result of alterations in the EPSPS amino acid coding sequence. These variants typically have a higher Ki for glyphosate than the wild-type EPSP enzyme which confers the glyphosate tolerant phenotype ‘Phenotype’ means the physical characteristics of the organism, as distinct from its genotype, which denotes its genetic make up. , but these variants are also characterised by a high Km for PEP which makes the enzyme kinetically less efficient.’
‘Class II epsps enzymes can be readily distinguished from Class I EPSPS’s by their inability to react with polyclonal antibodies prepared from Class I EPSPS enzymes under conditions where other Class I EPSPS enzymes would readily react with the Class I antibodies.’
‘[0012] Therefore, in one aspect, the present invention provides a new class of EPSP synthases that exhibit a low Km for phosphoenolpyruvate (PEP), a high Vmax/Km ratio, and a high Ki for glyphosate such that when introduced into a plant, the plant is made glyphosate tolerant such that the catalytic activity of the enzyme and plant metabolism are maintained in a substantially normal state. For the purposes of this discussion, a highly efficient EPSPS refers to its efficiency in the presence of glyphosate.’
‘14 5 A method of producing genetically transformed plants which are tolerant toward glyphosate herbicide, comprising the steps of: a) inserting into the genome of a plant cell a recombinant, double-stranded DNA molecule comprising: i) a promoter which functions in plant cells to cause the production of an RNA sequence, selected from the group consisting of CaMV35S and FMV35Spromoters, ii) a structural DNA sequence that causes the production of an RNA sequence which encodes a 5 fusion polypeptide comprising an amino terminal chloroplast transit peptide and a Class II EPSPS enzyme capable of reacting with antibodies raised against a Class II EPSPS enzyme selected from the group consisting of the enzymes of SEQ ID NO:3 and SEQ ID NO:5, iii) a 3 non-translated dna sequence which functions in plant cells to cause the addition of a stretch of polyadenyl nucleotides to the 3end of the RNA sequence being the NOS 3 polyadenylation sequence fragment where the promoter is heterologous with respect to the structural DNA sequence and adapted to cause sufficient expression of the fusion polypeptide to enhance the glyphosate tolerance of a plant cell transformed with said gene and where the fusion polypeptide has the amino acid sequence of SEQ ID NO: 15 fused through the common terminal methionine to the amino acid sequence of SEQ ID NO:3 in which serine at position 2 is replaced by leucine; b) obtaining a transformed plant cell; and c) regenerating from the transformed plant cell a genetically transformed plant which has increased tolerance to glyphosate herbicide.’ a) inserting into the genome of a plant cell a recombinant, double-stranded DNA molecule comprising: i) a promoter which functions in plant cells to cause the production of an RNA sequence, selected from the group consisting of CaMV35S and FMV35Spromoters, ii) a structural DNA sequence that causes the production of an RNA sequence which encodes a 5 fusion polypeptide comprising an amino terminal chloroplast transit peptide and a Class II EPSPS enzyme capable of reacting with antibodies raised against a Class II EPSPS enzyme selected from the group consisting of the enzymes of SEQ ID NO:3 and SEQ ID NO:5, iii) a 3 non-translated dna sequence which functions in plant cells to cause the addition of a stretch of polyadenyl nucleotides to the 3end of the RNA sequence being the NOS 3 polyadenylation sequence fragment b) obtaining a transformed plant cell; and c) regenerating from the transformed plant cell a genetically transformed plant which has increased tolerance to glyphosate herbicide.’
‘A glyphosate tolerant plant cell comprising a DNA molecule of Claims 8, 9, 12 or 134.’
‘ Mr Prescott QC and Mr Howe QC, for the defendants, submitted that the first step, as always, is to construe the claims in each patent. Here their construction is not in doubt. In 649 the process ends with the production of the father; in each of the others with the production of the master. They and they alone are the products obtained directly by means of the processes. Mr Prescott and Mr Howe accepted that a product can be further processed without losing its identity, in which event it remains the product obtained directly by means of the patented process. The question whether it has lost its identity depends on whether it no longer retains its essential characteristics. It is only for that purpose that a consideration of essential characteristics is appropriate. There is no free-standing "essential characteristics" test. Mr Prescott and Mr Howe further submitted that here there is no identity between the masters and the finished discs. Father, mother and son are each separate products with identities different from that of the master. However you look at it, the finished discs cannot properly be described as products obtained directly by means of the patented processes.’
‘There is no doubt that, where the matter alleged to amount to anticipation consists of written description, the interpretation of that description is, like the interpretation of any document, a question for the court assisted where necessary by evidence regarding the meaning of technical language. It was argued that the same applied to a photograph. I do not think so. Lawyers are expected to be experts in the use of the English language, but we are not experts in the reading or interpretation of photographs. The question is what the eye of the man with appropriate engineering skill and experience would see in the photograph, and that appears to me to be a matter for evidence. Where the evidence is contradictory, the judge must decide. But the judge ought not, in my opinion, to attempt to read or construe the photograph himself; he looks at the photograph in determining which of the explanations given by the witnesses appears to be most worthy of acceptance.’
‘Chemical bonds are formed by electrons, and the rearrangement or breakage of bonds requires the migration of electrons. In broad terms, reactive chemical groups can be said to function either as electrophiles or as nucleophiles. Electrophiles are electron-deficient substances that react with electron-rich substances; nucleophiles are electron-rich substances that react with electron deficient substances. The task of a catalyst often is to make a potentially reactive group more reactive by increasing its intrinsic electrophilic or nucleophilic character. In may cases the simplest way to do this is to add or remove a proton.’
‘Promoters which are known or are found to cause transcription of DNA in plant cells can be used in the present invention. Such promoters may be obtained from a variety of sources such as plants and plant DNA viruses and include, but are not limited to, the CaMV35S and FMV35S promoters and promoters isolated from plant genes such as ssRUBISCO genes.’
‘encoding a Class II EPSPS enzyme, said enzyme being an EPSPS enzyme having a Km for phosphoenolpyruvate (PEP) between 1-150 µM and a Ki(glyphosate)/Km(PEP) ratio between 3-500 which enzyme is capable of reacting with antibodies raised against a Class II EPSPS enzyme selected from the group consisting of the enzymes of SEQ ID NO:3 and SEO ID NO:5.’
‘Class II epsps enzymes can be readily distinguished from Class I EPSPS’s by their inability to react with polyclonal antibodies prepared from Class I EPSPS enzymes under conditions where other Class I EPSPS enzymes would readily react with the Class I antibodies.’
‘[0034] When crude extracts of CP4 and LBAA bacteria (50 µg protein) were probed using rabbit anti-EPSPS antibody (Padgette et al. 1987) to the Petunia EPSPS protein in a Western analysis, no positive signal could be detected, even with extended exposure times (Protein A-125I development system) and under conditions where the control EPSPS (Petunia EPSPS, 20 ng: a Class I EPSPS) was readily detected. The presence of EPSPS activity in these extracts was confirmed by enzyme assay. This surprising result, indicating a lack of similarity between the EPSPS’s from these bacterial isolates and those previously studied, coupled with the combination of a low Km for PEP and a high Ki for glyphosate, illustrates that these new EPSPS enzymes are different from known EPSPS enzymes (now referred to as Class I EPSPS).’
‘Apart from the, frankly desperate, suggestion that Zeneca scientists with substantial experience in running gels could not load the gels properly, Professor Lichtenstein’s evidence is merely that he would have liked to have watched everything to see if he could find fault.’
‘There are, we think, four steps which require to be taken in answering the jury question. The first is to identify the inventive concept embodied in the patent in suit. Thereafter, the court has to assume the mantle of the normally skilled but unimaginative addressee in the art at the priority date and to impute to him what was, at that date, common general knowledge in the art in question. The third step is to identify what, if any, differences exist between the matter [forming part of the state of the art] and the alleged invention. Finally, the court has to ask itself whether, viewed without any knowledge of the alleged invention, those differences constitute steps which would have been obvious to the skilled man or whether they require any degree of invention.’
‘Class II EPSPS enzymes often may be distinguished from Class I EPSPS’s by their inability to react with polyclonal antibodies prepared from Class I EPSPS enzymes under conditions where other Class I EPSPS enzymes would readily react with the Class I antibodies as well as the presence of certain unique regions of amino acid homology which are conserved in Class II EPSP synthases as discussed hereinafter.[my emphasis]’
‘We are interested in elucidating how PG2982 breaks down glyphosate and whether a “phosphonatase” enzyme similar to that characterised in B cereus by [reference omitted] is involved in the degradation. Assays for the degradation of glyphosate and Amph with undialyzed and dialyzed cell-free extracts have been unsuccessful…’
“Vorträge für Pflanzenzüchtung”
‘Further evidence for correct delivery of the bacterial EPSP synthase into chloroplasts comes from the observation that plants expressing the fusion 3 polypeptide display a distinctive glyphosate tolerance phenotype readily distinguishable from the one conferred by cytoplasmic compartmentation of the aroA encoded EPSP synthase. Transgenic tomato plants expressing fusion 3 were tested in a field trial in Yolo County, California, in summer 1988. Application of 1 kg/ha of glyphosate salt (Roundup) at the 2-3 true leaf stage, or at the 5 to 6 true leaf stage, caused no loss in tomato yield. ’
‘The recent intrigue about the genetic engineering of [glyphosate] resistance into plants has led to formulation of objectives for genetic alteration of EPSP synthases or identification of genes which code for [glyphosate] degradation. Another approach has been the search for EPSP synthase enzymes in nature that exhibit natural tolerance to inhibition by [glyphosate]. In this context, we noticed the apparent lack of sensitivity to inhibition by [glyphosate] displayed by EPSP synthase from Bacillus subtilis. The data reported here demonstrate that when EPSP synthase is characterized under what may otherwise be standard conditions of assay, the true sensitivity to inhibition by glyphosate may be masked. In particular, the cation requirements of EPSP synthases may be directly linked to sensitivity to [glyphosate]-promoted inhibition.’
‘If the cations contributed by the assay components are less than required for maximal activation, EPSP synthases of the B subtilis type would falsely appear to be relatively resistant to inhibition by [glyphosate] in vitro. Before assigning relative sensitivities of EPSP synthase enzymes to [glyphosate]-mediated inhibition, an evaluation of possible modulation of enzyme activity by monovalent cations seems essential. We also recommend the utilisation of Hepes/tetramethylammonium buffer plus the preparation of all substrates and [glyphosate] as tetramethylammonium salts if possible.’
‘EPSPS enzyme assays were performed using either the phosphate release or radioactive HPLC method, as previously described in Padgette et al. 1987, using 1mM [PEP] and 2mM shikimate-3-phosphate (S3P) substrate concentrations. For radioactive HPLC assays, 14C-PEP (Amersham) was utilized.’
‘The task of the Court is threefold: (1) to ascertain through the eyes of the skilled addressee what is disclosed, both explicitly and implicitly in the application. (2) To do the same in respect of the patent as granted. (3) To compare the two disclosures and decide whether any subject matter relevant to the invention has been added whether by deletion or addition. The comparison is strict in the sense that subject matter will be added unless such matter is clearly and unambiguously disclosed in the application either explicitly or implicitly.’