DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Priority
Applicant’s claim for the benefit of a prior-filed application no. 63/460,095 filed April 18th, 2023, and PCT/IL2024/050374 filed April 16th, 2024, under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged.
Thus, the earliest possible priority for the instant application is April 18th, 2023.
The later-filed application must be an application for a patent for an invention which is also disclosed in the prior application (the parent or original nonprovisional application or provisional
application). The disclosure of the invention in the parent application and in the later-filed application must be sufficient to comply with the requirements of 35 U.S.C. 112(a) or the first paragraph of pre-AIA 35 U.S.C. 112, except for the best mode requirement. See Transco Products, Inc. v. Performance Contracting, Inc., 38 F.3d 551, 32 USPQ2d 1077 (Fed. Cir. 1994).
The disclosures of the prior-filed applications, Application Nos. 63/460,095 and PCT/IL2024/050374, fail to provide adequate support or enablement in the manner provided by 35 U.S.C. 112(a) or pre-AIA 35 U.S.C. 112, first paragraph for one or more claims of this application. In particular, these applications do not include the SEQ ID NOs. of the instant application recited in claim 48.
Information Disclosure Statement
The information disclosure statements (IDSs) submitted on October 16th, 2025, and January 20th, 2026, were considered, initialed, and attached hereto. A signed copy of the list of references cited is included with this Office Action.
Status of Claims
Claims 38-57 filed October 16th, 2025, are pending and examined herein.
Claim Objections
Claim 38 objected to because of the following informalities: Claim 38 is objected to for the recitation of “…and the two or more genes encode at least one haloperoxidase enzyme and at least one other enzyme involved in bromoform production and not encoded by the plant,” in lines 5-6. This should be corrected for grammar to recite “…and the two or more genes encode at least one haloperoxidase enzyme and at least one other enzyme involved in bromoform production and are not encoded by the plant,” or some rephrasing for clarity. Appropriate correction is required.
Claim Interpretation
The term "pasture plant" or "forage plant" as recited in claim 53 is interpreted to refer to plants which are planted or seeded in grazing fields for animals to graze on, as stated in the instant specification [pg. 17, lns. 13-14].
The system for gene expression as recited in claim 38, and claims 37-57 depending therefrom, is interpreted to have practical utility in producing plants that can produce bromoform after transformation with the gene expression system and is thus not rejected as lacking utility under 35 USC § 101.
Claim 38 is interpreted to be a system for gene expression in a plant comprising two or more genes, wherein the first gene encodes one haloperoxidase and the other gene encodes another enzyme involved in bromoform production, both of which are not naturally encoded by the plant.
Claim Rejections - 35 USC § 112(a)
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Written Description
Claims 38-57 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
The instant specification describes:
Cloning of bromoform producing genes and expression of plasmids containing said genes in Arabidopsis, tobacco, Alfalfa, and Brachaiaria.
RT-PCR analysis of transformed tobacco plants using specific primers to identify the integration and transcription of the bromoform-producing genes per transgenic plant.
A procedure for bromoform production testing.
The instant specification does not describe:
That the system for gene expression in a plant is able to function in producing bromoform in any plant.
The claims are broadly drawn to a system for gene expression in any plant, comprising one or more recombinant nucleic acid molecules comprising one or nucleotide sequences comprising two or more genes. Each gene is operably linked to a promoter for expression in the plant. The gene(s) encode at least one haloperoxidase enzyme and at least one other enzyme involved in bromoform production, and are not encoded by the plant. The claims further limit to require a genetically engineered bromoform-producing plant or plant part comprising the system. Under the claim interpretation detailed above, the claims are interpreted to have specific utility as the system for gene expression in a plant appears to engineer the plant to produce bromoform.
The Applicant describes several exemplary plant species transformed with the gene expression system and states that “it is expected that plants capable of growing on the selectable markers, confirming successful transformation with bromoform-producing genes, will be able to produce bromoform” [Example 2]. The Applicant further states that the transformed plants are tested for bromoform production three-weeks post germination, as described in Example 7. Example 7 provides a procedure for bromoform production testing by GC/MS analysis, but no data is furnished to support the claim that the system for gene expression in a plant functions to produce bromoform when comprised within a plant or plant part.
The only data shown is displayed in Fig. 1A-C, which shows a PCR analysis of transformed tobacco plants using primers to identify the integration and transcription of the genes. Although the figures show that the genes may be expressed, confirmed transcription does not necessitate that the plant will function as intended. The mRNA still needs to be translated into a protein, which requires additional challenges not considered in the instant application. This includes factors such as protein folding, post-translational changes, cell location, and accidental gene silencing. For example, specialized metabolic pathways differ in plants across phylogeny and heterologous introduction of multiple biosynthetic genes in a plant requires optimization of expression levels and accumulation (Calgaro-Kozina, A. et al. 2020. “Engineering Plant Synthetic Pathways for the Biosynthesis of Novel Antifungals.” ACS Cent. Sci. 6:1394-1400) [pg. 1398, col. 1, ¶1]. Further, expression of core pathway genes can result in different abundance from when they are expressed individually and enzymes may competitively inhibit one another along the pathway [pg. 1396, col. 2, ¶1].
The art demonstrates significant unpredictability in the structure function relationship of genetically engineering biosynthetic pathways into plants to produce a compound and cannot remedy the deficiencies of the instant specification. The Applicant has not reduced a sufficient number of species to practice to support the claim of the gene expression system having adequate structure to perform the function. Undue experimentation would be required for one of ordinary skill in the art to validate and test the genes claimed in the instant claims to result in a genetically engineered plant that is capable of producing bromoform.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
Claims 38-41, 46, and 49-57 are rejected under 35 U.S.C. 103 as being unpatentable over Peacock, C. et al. International Publication No. WO 2024110672 A1. Effectively filed 11/25/2022. (see, IDS filed 01/20/2026), in view of Thapa, H. et al. (2020). “Genetic and Biochemical Reconstitution of Bromoform Biosynthesis in Asparagopsis Lends Insights into Seaweed Reactive Oxygen Species Enzymology.” ACS Chem Biol. 15(6):1662-1670. (see, IDS filed 10/16/2025).
Claim 38 recites a system for gene expression in a plant, comprising one or more recombinant nucleic acid molecules which comprise one or more nucleotide sequences together comprising two or more genes, wherein: each gene is operably linked to a promoter for expression in the plant; and the two or more genes encode at least one haloperoxidase enzyme and at least one other enzyme involved in bromoform production and not encoded by the plant.
Claim 39 recites the system of claim 38, wherein at least one of the two or more genes is from an organism selected from the group consisting of Asparagopsis taxiformis.
Claim 40 recites the system of claim 38, wherein at least two of the two or more genes are from a non-plant organism.
Claim 41 recites the system of claim 38, wherein the at least one other enzyme comprises an NAD(P)H Oxidase (NOX).
Claim 46 recites the system of claim 38, wherein the two or more genes are selected from the group consisting of Mbb 1 (A. taxiformis), Mbb4 (A. taxiformis).
Claim 49 recites the system of claim 38, wherein at least one of the promoters is a constitutive promoter.
Claim 50 recites the system of claim 38, wherein at least one of the promoters is an inducible promoter.
Claim 51 recites the system of claim 50, wherein the inducible promoter is a tissue-specific promoter, activated in plant organs which develop late in the plant life cycle, such as leaves or flowers.
Claim 52 recites the system of claim 38, wherein at least one of the two or more genes further encodes a chloroplast transit peptide for targeting the enzymes to a chloroplast.
Claim 53 recites the system of claim 38, wherein the plant is a pasture plant or a fodder plant.
Claim 54 recites the system of claim 38, wherein the plant is selected from the group consisting of millet.
Claim 55 recites a composition comprising the system of claim 38.
Claim 56 recites a genetically engineered bromoform-producing plant or plant part, comprising the system of claim 38.
Claim 57 recites the bromoform-producing plant or plant part of claim 56, wherein the plant part is selected from the group consisting of a leaf.
Regarding claim 38, Peacock teaches that ruminant livestock emit methane, a potent greenhouse gas [¶2]. Various macro algae have been shown to reduce methane emissions when fed to ruminants, with red macro alga Asparagopsis sp. being shown to be highly effective [¶54]. These species produce halogenated compounds, including bromoform (CH8r3) and polybromomethanes such as dibromomethane and chlorodibromomethane, which are thought to be synthesised by haloperoxidases. Peacock teaches a nucleic acid construct for inducing expression of a polypeptide in a plant cell (i.e., a system for gene expression in a plant, comprising one or more recombinant nucleic acid molecules which comprising one or more nucleotide sequences together), the nucleic acid construct comprising a coding sequence encoding a polypeptide comprising a vandate-dependent haloperoxidase (VHPO) (i.e., at least one haloperoxidase) operably linked to one or more regulatory elements (i.e., wherein each gene is operably linked to a promoter for expression in the plant) [¶13]. This provides an improved way to reduce methane production and to improve feed efficiency [¶12].
Peacock teaches that the VHPO can be Mbb1, Mbb3, and/or Mbb4 of Asparagopsis taxiformis [¶19]. Peacock teaches expression of AtMbb1, CcMbb3 and AaMbb4-2 in corn protoplasts [¶100; Example 1] and teaches expression of Mbb1 in Arabidopsis plants using the same signal peptides used in transient expression in the corn protoplasts [¶110; Example 2]. Peacock further teaches that the modified plants or plant cells can be modified to further produce or provide substrates, cofactors, and/or other compounds or conditions necessary for the VHPO to function [¶74]. For example, the plants or plant cells can be modified to produce further ROS, such as by modification to produce a protein that produces or increases production of ROS.
Although Peacock does not explicitly provide a working example in which the system for gene expression in a plant comprises two genes, wherein the second gene encodes an enzyme involved in bromoform production, the biosynthetic pathway for bromoform was reconstituted prior to the filing date of the instant application as taught by Thapa.
Thapa teaches the molecular identities of the enzymes participating in hydrogen peroxide and bromoform production and reconstitute their respective biochemical activities [pg. 4, ¶1]. Thapa teaches that the requirement for vanadate in bromoform production lends support that VHPOs, halogenating enzymes widely detected in other seaweeds, are involved in bromoform biosynthesis [pg. 5, ¶3] and undertook a transcriptome sequencing and mining approach [pg. 5, ¶4]. Mbb1, Mbb3, and Mbb4 identified in the Guam transcriptome were cloned from cDNA into plasmid vectors for expression in Escherichia coli [pg. 6, ¶2]. Thapa teaches that the presence of different haloperoxidases allows them to access different hydrocarbon substrate pools, or, that they are trafficked to different subcellular locations for bromoform production [pg. 10, ¶1]. This suggests that expression of multiple haloperoxidases, or enzymes involved in bromoform production, would lead to higher bromoform production.
Given that Peacock teaches a system for gene expression in a plant comprising one or more recombinant nucleic acid molecules comprising at least one haloperoxidase gene for bromoform production, and suggests the addition of other genes related to bromoform production; and given that Thapa teaches the bromoform biosynthetic pathway, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to express an additional gene involved in bromoform production because the biosynthetic pathway for bromoform was reconstituted prior to the filing date of the instant application. As taught by both Peacock, the development of plants that can produce bromoform, which binds to reduced vitamin B12, inhibiting methane formation, as a food source for ruminant livestock can reduce methane production [¶53]. One would have been motivated to include another gene involved in bromoform production, such as other haloperoxidase genes, as Thapa teaches the importance of the presence of different haloperoxidases (Mbb1-4) in improving bromoform biosynthesis. One would have reasonable expectation of success given that Peacock teaches an expression system in plants and corn protoplasts and Thapa teaches expression in E. coli, demonstrating the methods of expression of bromoform-related genes previously known in the art.
Regarding claim 39, Peacock and Thapa teach that the haloperoxidase genes used in the expression constructs are isolated from Asparagopsis taxiformis (i.e., wherein at least one of the two or more genes is from an organism selected from the group consisting of Asparagopsis taxiformis) [Peacock, ¶18; Thapa, pg. 5, ¶4].
Regarding claim 40, as detailed in the rejection of claim 38 above, Peacock and Thapa teach expression of various haloperoxidase enzymes which are isolated from species such as Asparagopsis taxiformis, an algae organism (i.e., wherein at least two of the two or more genes are from a non-plant organism) [Peacock, ¶18; Thapa, pg. 5, ¶4].
Regarding claim 41, Peacock further teaches that the modified plants or plant cells can be modified to further produce or provide substrates, cofactors, and/or other compounds or conditions necessary for the VHPO to function [¶74]. For example, the plants or plant cells can be modified to produce further ROS, such as by modification to produce a protein that produces or increases production of ROS. Such a protein may be an NADPH_oxidase, for example, A. taxiformis NADPH_oxidase Mbb2 (i.e., wherein the at least one other enzyme comprises an NAD(P)H Oxidase). Peacock teaches that haloperoxidases catalyse the conversion of halide anions to hypohalous acid with hydrogen peroxide (H2O2) acting as the oxidant [¶54]. The requirement for H2O2 links synthesis to the enzymatic production of reactive oxygen species (ROS).
Further, Thapa teaches that the first hydrogen peroxide-producing NOX serves as an initiation point to determine, at the molecular level, the ROS enzymology in seaweed and the response of these enzymes to biotic and abiotic stressors [pg. 10, ¶2]. Biosynthesis of halogenated molecules in seaweeds is inextricably linked to reactive oxygen species (ROS) signaling as hydrogen peroxide serves as a substrate for haloperoxidase enzymes that participate in the construction these halogenated molecules [Abstract].
It would have been prima facie obvious to one of ordinary skill in the art at the time of filing to modify the system for gene expression as taught by Peacock to include an NADPH oxidase, such as Mbb2 from A. taxiformis, which is involved in bromoform production, as taught by Thapa. One would have been motivated to do so given that increased production of ROS leads to generation of H2O2, which acts as an essential substrate for VHPO to utilize bromide, and thus drive biosynthesis of bromoform. Peacock provides the suggestion to include further modifications to the engineered plant, such as further producing ROS with NADPH_oxidase Mbb2 from A. taxiformis. One would have reasonable expectation of success given that other Mbb enzymes have been successfully expressed in the defined systems for gene expression in plants and given that Mbb2/NOX was previously identified in the biosynthetic pathway.
Regarding claim 46, Thapa and Peacock teach expression of Mbb1 and Mbb4 haloperoxidase enzymes which are isolated from Asparagopsis taxiformis (i.e., wherein the two or more genes are selected from the group consisting of Mbb1 (A. taxiformis) and Mbb4 (A. taxiformis) [Peacock, ¶18; Thapa, pg. 5, ¶4].
Regarding claim 49-50, Peacock teaches that the promoter may be a constitutive or inducible promoter (i.e., wherein at least one of the promoters is a constitutive promoter/inducible promoter) [¶25].
Regarding claim 51, Peacock teaches that the plant gene inducible promoters include those for tissue-specific expression, including those that are regulated in response to developmental signals (i.e., wherein the inducible promoter is a tissue-specific promoter, activated in plant organs which develop late in the plant life cycle, such as leaves or flowers) [¶69-70].
Regarding claim 52, Peacock teaches that since the VHPO proteins are peroxidases, they need H2O2, and potentially carbon feedstocks, in order to function, the availability of these substrates varies within cells and may be more abundant in chloroplasts, since chloroplasts are particularly involved in the production and/or removal or ROS [¶76]. Peacock teaches that targeting to specific organelles can help to protect the cell from any deleterious effects [¶77]. Peacock teaches the addition of a chloroplast transit peptide in the construct design for targeting (i.e., wherein at least one of the two or more genes further encodes a chloroplast transit peptide for targeting the enzymes to a chloroplast) [¶98].
Regarding claims 53-54, Peacock teaches that the nucleic acid construct can be introduced into corn, millet, sorghum, or pasture grasses (i.e., wherein the plant is a pasture plant or a fodder plant; wherein the plant is selected from the group consisting of millet) [claim 26b].
Regarding claims 55-57, Peacock teaches expression of the gene expression system in Arabidopsis, which resulted in the production of bromoform (i.e., a composition comprising the system of claim 38; a genetically engineered bromoform-producing plant or plant part comprising the system of claim 38) [Example 5]. Peacock teaches in vivo production of bromoform by transgenic Arabidopsis lines, teaching that seedlings or rosette leaf tissue for Arabidopsis were tested for production of bromoform (i.e., the bromoform-producing plant or plant part of claim 56, wherein the plant part is selected from the group consisting of a leaf) [¶135].
Thus, Peacock and Thapa combined render obvious the gene system and compositions and plants thereof.
Claims 42-45 and 47 are rejected under 35 U.S.C. 103 as being unpatentable over Peacock and Thapa as applied to claim 38 above, and further in view of Gengenbach, B. et al. “Methods for expressing a maize acetyl CoA carboxylase gene in host cells and encoded protein produced thereby.” US. Patent Application Publication No. US 6146867 A. Published 11/14/2000.
Claim 42 recites the system of claims 38, wherein the at least one other enzyme comprises at least one enzyme involved in fatty acid synthesis.
Claim 43 recites the system of claim 42, wherein the at least one enzyme involved in fatty acid synthesis is selected from the group consisting of Beta-ketoacyl-[acyl-carrier-protein] synthase III (FabH), phosphopantetheinyl transferase (sfp), acetyl-CoA carboxylase (ACCase), and malonyl CoA synthetase (MatB).
Claim 44 recites the system of claim 43, wherein the at least one other enzyme comprises NOX,and at least one enzyme involved in fatty acid synthesis selected from the group consisting of FabH, sfp, ACCase, and MatB.
Claim 45 recites the system of claim 44, wherein the at least one other enzyme comprises NOX,FabH, sfp, and ACCase.
Claim 47 recites the system of claim 46, wherein the two or more genes comprise Mbbl, Mbb2,Mbb3, Mbb4, and at least one gene selected from the group consisting of FaBH, Sfp, ACCase, and MatB.
Regarding claims 42-43, as detailed above, Peacock and Thapa render obvious the system of claim 38. Thapa teaches that bromoform can be derived from fatty acid biosynthesis. Thapa teaches that fatty acid biosynthetic intermediates can served as substrate for bromoform production, purifying enzymes involved in fatty acid synthesis, such as FabH, Sfp, ACCase, and MatB [pg. 6, ¶4-5]. Thapa teaches that it is likely that the primary hydrocarbon substrate(s) employed by Asparagopsis for bromoform production are endogenously produced.
Thapa does not explicitly teach that the system for gene expression includes a recombinant nucleic acid molecule, wherein the at least one other enzyme comprises at least one enzyme involved in fatty acid synthesis, however, recombinant nucleic acid molecules encoding genes involved in fatty acid biosynthesis are well known in the art.
For example, Gengenbach teaches that an important property of ACCase is the central role it plays in fatty acid synthesis and accumulation in plants and seeds [¶11]. Available evidence supports the idea that ACCase activity is the rate-limiting step for de novo fatty acid synthesis in plants. Gengenbach teaches that ACCase catalyzes the formation of malonyl-CoA from acetyl-CoA and bicarbonate in animal, plant, and bacterial cells [¶3]. Malonyl-CoA is an essential substrate for (i) de novo fatty acid (FA) synthesis, (ii) fatty acid elongation, (iii) synthesis of secondary metabolites such as flavonioids and anthocyaninis, and (iv) malonylation of some amino acids and secondary metabolites. Synthesis of malonyl-CoA is the first committed step of flavonoid and fatty acid synthesis and thus current evidence suggests that ACCase catalyzes the rate-limiting step of fatty acid synthesis.
Gengenbach teaches a method introducing and expressing a plant acetyl CoA carboxylase (ACCase) gene in plant cells (wherein the at least one other enzyme comprises at least one enzyme involved in fatty acid synthesis; wherein the at least one enzyme involved in fatty acid synthesis is selected from the group consisting of ACCase) [Abstract]. The method includes the steps of introducing an expression cassette encoding a plant ACCase operably linked to a promoter functional in plant cells, into the cells of a plant tissue and expressing the plant ACCase gene. The expression cassette can also be introduced into other host cells to increase yield of a plant ACCase enzyme.
Given that Thapa teaches that fatty acid biosynthetic intermediates can serve as substrates for bromoform production and are endogenously produced by Asparagopsis for bromoform production, and given that expression of fatty acid intermediates was known in the art at the time of filing, it would have been prima facie obvious to one of ordinary skill to have included a recombinant nucleic acid molecule comprising a plant ACCase operably linked to a promoter for expression in the plant, as taught by Gengenbach. One would have been motivated to increase fatty acid biosynthetic intermediate availability as a substrate for increased bromoform production, especially given that Asparagopsis endogenously produce substrate, indicating that overexpression of potential substrates may aid in bromoform production. One would have reasonable expectation of success given that Gengenbach explicitly teaches a recombinant nucleic acid molecule effective for expressing at least one enzyme involved in fatty acid synthesis.
Regarding claim 44, the arrangement of the recombinant acid molecules within the system for gene expression would be merely routine optimization given that all elements are present in the prior art. As detailed above, Peacock and Thapa teach the potential for an additional enzyme to be NOX, Thapa teaches the importance of endogenous fatty acid biosynthesis to enhance substrate availability for bromoform production, and Gengenbach teaches overexpression of a known enzyme involved in fatty acid biosynthesis, ACCase (i.e., wherein the at least one other enzyme comprises NOX, at the at least one other enzyme involved in fatty acid synthesis is selected from the group consisting of ACCase).
Regarding claim 45, Thapa teaches the importance of NOX, FabH, Sfp, and ACCase in fatty acid biosynthesis and teaches characterization of these enzymes. Gengenbach teaches overexpression of ACCase to enhance fatty acid biosynthesis (i.e., wherein the at least one other enzyme comprises NOX, FabH, sfp, and ACCase). Given that a method for overexpression of one fatty acid biosynthesis gene was known and the remaining genes characterized with their functionality relating to bromoform production suggested in the prior art at the time of filing, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to try to expression other fatty acid biosynthesis genes in plants to increase fatty acid biosynthesis for a substrate of bromoform production. These genes are known alternatives or additives to the methodology provided for ACCase, are there are a finite number of genes available within the fatty acid biosynthetic pathway to try. One would have reasonable expectation of success in overexpressing such genes, given that Gengenbach provides clear methodology and Thapa teaches motivation to enhance a substrate for bromoform production. There was a clear pressure to solve the problem of methane production by utilizing such gene expression systems with fatty acid biosynthetic genes to inhibit methane production with bromoform.
Regarding claim 47, Thapa and Peacock teach expression of Mbb1 and Mbb4 haloperoxidase enzymes which are isolated from Asparagopsis taxiformis (i.e., wherein the two or more genes comprise Mbb1, Mbb4) [Peacock, ¶18; Thapa, pg. 5, ¶4]. As detailed above, Gengenbach teaches a gene expression construct in a plant expressing ACCase for increased fatty acid biosynthesis, which can enhance bromoform production, as taught by Thapa (i.e., and at least one gene selected from the group consisting of ACCase).
Claim 48 is rejected under 35 U.S.C. 103 as being unpatentable over Peacock, Thapa, and Gengenbach as applied to claim 44 above, and further in view of Ayliffe, M. et al. “Methods.” US. Publication No. US 20250051805 A1. Effectively filed 06/24/2022.
Claim 48 recites the system of claim 44, wherein: the haloperoxidase has at least 70% sequence identity to SEQ ID NO: 1 (Mbb 1).
Regarding claim 48, as detailed above, Peacock, Thapa, and Gengenbach render obvious the system of claim 44, wherein Peacock and Thapa teach a system for gene expression in a plant comprising one or more recombinant nucleic acid molecules, wherein one gene encodes a haloperoxidase enzyme and the other encodes at least one enzyme involved in bromoform production. Gengenbach further teaches an expression construct for increasing fatty acid biosynthesis, which is a substrate for bromoform, as taught by Thapa.
Peacock, Thapa and Gengenbach do not explicitly teach wherein the haloperoxidase has at least 70% identity to SEQ ID NO: 1. However, Ayliffe teaches methods of increasing bromoform production [Abstract]. Ayliffe teaches genes encoding VHPO enzymes cloned from a number of algal and bacterial species and shown to produce bromoform under appropriate conditions when introduced into numerous organisms including Escherichia coli, Saccharomyces cerevisiae and Pichia pastoris. Ayliffe teaches a nucleic acid construct which comprises appropriate elements that allow the molecule to be incorporated into the genome, or the construct is placed in an appropriate vector which can be incorporated into a chromosome of a plant cell [¶267]. Ayliffe teaches expression of Asparagopsis taxiformis VHPO genes in transgenic plants, including Arabidopsis [Example 8].
Ayliffe further teaches Asparagopsis taxiformis vanadium-dependent haloperoxidase set forth in SEQ ID NO: 2 with 97.8% identity to SEQ ID NO: 1 of the instant application (i.e., wherein the haloperoxidase has at least 70% identity to SEQ ID NO: 1 (Mbb1) [¶109] (see, alignment below). Given that Peacock, Thapa, and Gengenbach render obvious the system of claim 44 and teach expression of an Asparagopsis taxiformis Mbb1 haloperoxidase in the gene system, and given that Ayliffe teaches a vanadium-dependent haloperoxidase in Asparagopsis taxiformis, it would have been prima facie obvious to one of ordinary skill in the art at the time of filing to use a known haloperoxidase that had already been successfully expressed in a plant for the production of bromoform. One would be motivated given that Ayliffe has the same methodology and motivation of increasing bromoform production, and one would have expectation of success given that the methodology was known at the time of filing.
Query Match 97.8%; Score 3022; Length 590;
Best Local Similarity 97.5%;
Matches 575; Conservative 8; Mismatches 7; Indels 0; Gaps 0;
Qy 1 MTDTQNPNRAEVAFKVRVSAAELARARGSPAHLSNNSESRFRNPDGTRSLLANFTKGLPH 60
|||||||||||||||||||||||||||||| |||||||||||||||||||||||||||||
Db 1 MTDTQNPNRAEVAFKVRVSAAELARARGSPTHLSNNSESRFRNPDGTRSLLANFTKGLPH 60
Qy 61 IKETALVESAIDYDRFVRAIDSGDPRDFADLPLGPQGVEPRFTSGIASDPEVGTRAWESG 120
||||||| |||||| |||||||||||||||||||||||||||||||||||||||||||||
Db 61 IKETALVVSAIDYDNFVRAIDSGDPRDFADLPLGPQGVEPRFTSGIASDPEVGTRAWESG 120
Qy 121 GAGLVFDLEGPDAQAVTMPPAPELDSDELVAEVTECYWMSLLRDVPFPTFESNSHIQAAA 180
|||||||||||||||||||||||||||||| |||||||||||||||||||||: |||:||
Db 121 GAGLVFDLEGPDAQAVTMPPAPELDSDELVTEVTECYWMSLLRDVPFPTFESDPHIQSAA 180
Qy 181 ESINNTQWIKFKNNPPAHLTAAERSRLRGPVTTANVFRGITPGDEVGPYLSQFLLVGTTG 240
||||||||||||::||||||||||||||||:|||||||||||||||||||||||||||||
Db 181 ESINNTQWIKFKDSPPAHLTAAERSRLRGPITTANVFRGITPGDEVGPYLSQFLLVGTTG 240
Qy 241 IANGNEVGDGFIQYGGMRMDQRVRVAKPHIDYMTTFGAYLDVQNAANVSGRELYKEEEPR 300
||||||||||||||||||||||||||| |||||||||||||||||||||||||| |||||
Db 241 IANGNEVGDGFIQYGGMRMDQRVRVAKSHIDYMTTFGAYLDVQNAANVSGRELYNEEEPR 300
Qy 301 FRFIHTPRDLATYVHFDALYQAYLNACIILLDIGAPFDSGIPFQLDNDIDKQQGFATFGG 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 FRFIHTPRDLATYVHFDALYQAYLNACIILLDIGAPFDSGIPFQLDNDIDKQQGFATFGG 360
Qy 361 PHILSLVTEVATRALKAVRFQKFNVHRRLRPEAIGARVDRYCATKAPEFAGAAKLSEALD 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 PHILSLVTEVATRALKAVRFQKFNVHRRLRPEAIGARVDRYCATKAPEFAGAAKLSEALD 420
Qy 421 KELLQKVHDHNKKQNLLSDRGNPRANDFNPDGEVSEGNLLMPMAFPEGSPMHPAYGAGHA 480
|:||||||||||||||||||||||||||||||:|||||||||||||||||||||||||||
Db 421 KKLLQKVHDHNKKQNLLSDRGNPRANDFNPDGDVSEGNLLMPMAFPEGSPMHPAYGAGHA 480
Qy 481 TVAGACVTVLKAFFDGGYRLPFCYITDEDGTGLQAVEIDEPLTVDGELNKICSNISIGRN 540
||||||||||||||||||||||||||:|||||||||||||||||||||||||||||||||
Db 481 TVAGACVTVLKAFFDGGYRLPFCYITNEDGTGLQAVEIDEPLTVDGELNKICSNISIGRN 540
Qy 541 WAGVHYFTDYIESIRIGEEIAIGILQEQKLTFSENFSMTLNKFDGSTIRI 590
||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 WAGVHYFTDYIESIRIGEEIAIGILQEQKLTFSENFSMTLNKFDGSTIRI 590
Conclusion
No claims allowed.
Contact Information
Any inquiry concerning this communication or earlier communications from the examiner should be directed to EMILY K. JOHNSON whose telephone number is (571)272-5761. The examiner can normally be reached Monday - Friday 7:30 am - 5:00 pm.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Bratislav Stankovic can be reached at 571-270-0305. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/EMILY K JOHNSON/Examiner, Art Unit 1662
/BRATISLAV STANKOVIC/Supervisory Patent Examiner, Art Units 1661 & 1662
