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 .
DETAILED ACTION
Claims 1-20 are pending. Claims 1-16 are withdrawn. Claims 17-20 are under examination.
Election/Restrictions
Applicant’s election without traverse of Group III, claims 17-20, in the reply filed on 11/24/2025 is acknowledged.
Claims 1-16 withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 11/24/2025.
Claim Objections
Claims 17, 19, and 20 are objected to because of the following informalities: the claims contain grammatical errors and convoluted language.
Regarding claim 17, it is apparent that GPGQQ AAAAA GPGQQ GPGQQ GPGQQ GPGEQ GPGSG is the amino acid sequence of SEQ ID NO:1 and GPGQQ AAAAA AAAAA GPGQQ GPGQQ GPGEQ GPGSG is the amino acid sequence of SEQ ID NO:2. Therefore, the second wherein clause of claim 17 may be amended to recite: wherein the gene fragments encode the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:2.
Claim 19 may be amended to recite: The bacterium according to claim 17, wherein the one or more genes encoding silk proteins are downstream of a tac promoter and a lac operon and are inserted at an attTn7 site of the Pseudomonas bacterial chromosome.
Claim 20 may be amended to recite: The bacterium according to claim 17, wherein the one or more genes encoding silk proteins are within a recombinant plasmid,
wherein the recombinant plasmid is a pSEVA plasmid comprising an isopropyl 13-D-1-thiogalactopyranoside-inducible promoter.
Appropriate correction is required.
Claim Rejections - 35 USC § 112(b)
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 17-20 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 17 recites a recombinant bacterium including a Pseudomonas sp. bacterium. A bacterium cannot include or comprise another bacterium. Therefore, the claim is considered indefinite as it is unclear if the claim means that the recombinant bacterium must be a Pseudomonas sp., or if the claim means something else.
Claim 17 further recites: hydrocarbyl groups including between about 5 and about 32 carbon atoms; and gene fragments including a construction having between about 2 fragment and about 64 repeated fragments. The term “about” is a term of approximation which renders the claim indefinite. The range that is encompassed by the term “about” is not defined by the claim, and the specification does not provide the upper and lower limits of the range encompassed by the term “about”. One of ordinary skill in the art would not be reasonably be apprised of the scope of the invention. It is therefore unclear how many carbon atoms and how many repeated gene fragments, or non-repeated gene fragments, are within approximation as required by the claim.
Claim 17 further recites: wherein the one or more genes comprise gene fragments including a construction having between about 2 fragment and about 64 repeated fragments, but does not recite a sequence or reference a SEQ ID NO of said fragments. It is unclear how many/what nucleotides make up a “fragment”, therefore, it is unclear what sequences are to be repeated.
Claim 17 lastly recites: wherein the gene fragments code for GPGQQ AAAAA GPGQQ GPGQQ GPGQQ GPGEQ GPGSG (SEQ ID NO:1), GPGQQ AAAAA AAAAA GPGQQ GPGQQ GPGEQ GPGSG (SEQ ID NO:2), or combinations thereof. It is unclear which amino acids of SEQ ID NO:1 and SEQ ID NO:2 are required to form “combinations thereof”, i.e., it is unclear how amino acids from SEQ ID NO:1 and how amino acids from SEQ ID NO:2 may be combined to generate a combination of SEQ ID NO:1 and 2, as required by the claim. Therefore, the scope of the claim is unclear.
Claims 18-20 are also rejected as they depend from claim 1.
Claim 18 recites the bacterium, i.e., singular form, of claim 17, which is a narrow limitation, followed by a broad limitation, wherein the Pseudomonas sp. bacteria, i.e., plural form, includes P. aeruginosa RR1, P. aeruginosa NCIMB 14923, P. oleovorans, P. putida GPol, or combinations thereof. Because the claim recites bacterium and bacteria, as well as “combinations thereof”, it is unclear if the claim means that: a) the recombinant bacterium may comprise genetic material from any one of the recited Pseudomonas sp., b) that multiple of the recited Pseudomonas sp. may be present in a bacterial mixture, or c) that the recombinant Pseudomonas bacterium may be selected from any of P. aeruginosa RR1, P. aeruginosa NCIMB 14923, P. oleovorans, or P. putida GPol. One of ordinary skill in the art would not recognize what is required by the claim.
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.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Using BRI, it is interpreted that any silk protein that comprises a combination of the amino acids of SEQ ID NO:1 and SEQ ID NO:2 meets the limitation of “combinations thereof” as required by the claim.
Claims 17 and 18 are rejected under 35 U.S.C. 103 as being unpatentable over Guznik et al., (Conversion of post consumer polyethylene to the biodegradable polymer polyhydroxyalkanoate. Appl Microbiol Biotechnol. 2014. 98:4223–4232; cited in IDS filed 08/15/2023), in view of Joshi et al., (US2020/0291078A1).
Regarding claim 17, Guzik teaches Pseudomonas species capable of utilizing polyethylene pyrolysis wax hydrocarbons (Fig. 2; Table 2), i.e., pyrolysis products of polyethylene.
Guzik does not teach that the Pseudomonas sp. encode silk protein genes or a combination of SEQ ID NOs:1 and 2.
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However, Joshi teaches a recombinant bacterium expressing a gene encoding a silk fibrin domain (SEQ ID NO:3). SEQ ID NO:3 of Joshi shares 38.7% sequence identity with the combination of SEQ ID NOs:1 and 2 (see alignment below).
It would have been obvious to one of ordinary skill in the art, prior to the effective filing date of the claimed invention, to have engineered the Pseudomonas bacteria of Guznik to express and secrete the recombinant silk protein of Joshi. One of ordinary skill in the art would have been motivated to do so because Joshi teaches that the large-scale production of spider silk using spiders themselves is difficult since spiders are venomous and cannibalistic, therefore, spider silk production in bacteria is being explored (para. [0004 and 0005]). One of ordinary skill in the art would have had a reasonable expectation of success because Guznik and Joshi are in the same field of endeavor of industrial microbiology.
Regarding claim 18, Guzik teaches that P. oleovorans was a strain capable of utilizing polyethylene pyrolysis wax as its sole source of carbon (Table 2).
Claim 19 is rejected under 35 U.S.C. 103 as being unpatentable over Guznik et al., (Conversion of post consumer polyethylene to the biodegradable polymer polyhydroxyalkanoate. Appl Microbiol Biotechnol. 2014. 98:4223–4232) and Joshi et al., (US2020/0291078A1) as applied to claim 17 above, and further in view of Choi (mini-Tn7 insertion in bacteria with single attTn7 sites: example Pseudomonas aeruginosa. NATURE PROTOCOLS. Vol. 1 No. 1. 2006).
See discussion of Guznik above, which is incorporated into this rejection as well.
Joshi teaches that an inducible promoter may be the tac promoter or the lac operon promoter (para. [0180]). It would have been obvious to one of ordinary skill in the art that both promoters may be used to control expression of the genes encoding the silk proteins as both promoters are induced by IPTG.
Guznik and Joshi does not teach that the genes are inserted at an attTn7 site of the bacterium.
However, Choi teaches inserting genes at the attTn7 (Abstract).
It would have been obvious to one of ordinary skill in the art, prior to the effective filing date of the claimed invention, to insert genes encoding silk proteins downstream of tac and lac operon promoters, as taught by Joshi, at an attTn7 site as taught by Choi, of the Pseudomonas sp. taught by Guznik. One of ordinary skill in the art would have been motivated to insert the genes into the bacterial chromosome, i.e., the attTn7 site, because Choi teaches that plasmid cloning systems have several drawbacks such a necessity for continued selection and limited host range (Introduction, para. 1). One of ordinary skill in the art would have had a reasonable expectation of success because Joshi and Choi are in the same field of endeavor of bacterial genetic engineering.
Claim 20 is rejected under 35 U.S.C. 103 as being unpatentable over Guznik et al., (Conversion of post consumer polyethylene to the biodegradable polymer polyhydroxyalkanoate. Appl Microbiol Biotechnol. 2014. 98:4223–4232) and Joshi et al., (US2020/0291078A1) as applied to claim 17 above, and further in view of Aparicio et al., (CRISPR/Cas9-Based Counterseletion Boosts Recombineering Efficiency in Pseudomonas putida. Biotechnol. J. 2018, 13)
See discussion of Guznik above, which is incorporated into this rejection as well.
Joshi teaches that the recombinant silk protein is in a plasmid (para. [0054]), and teaches an IPTG-inducible promoter may be used for the invention (para. [0180]). Joshi teaches exemplary plasmid vectors (para. [0177]), but does not teach a pSEVA plasmid.
However, Aparicio teaches a pSEVA plasmid (Abstract), i.e., a plasmid containing a pSEVA backbone.
It would have been obvious to one of ordinary skill in the art, prior to the effective filing date of the claimed invention, to substitute the pSEVA plasmid taught by Aparicio for the plasmids taught by Joshi, to genetically modify the Pseudomonas bacteria taught by Guznik. One of ordinary skill in the art would have had a reasonable expectation of success because substituting one expression vector for another is well within the knowledge of one of ordinary skill, and the results of protein expression are reasonably predictable.
Conclusion
Any inquiry concerning this communication or earlier communications from the examiner should be directed to RACHEL EMILY MARTIN whose telephone number is (703)756-1416. The examiner can normally be reached M-Th 8:30-16:00, F 8:30-10:00 EST.
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/LOUISE W HUMPHREY/Supervisory Patent Examiner, Art Unit 1657
/RACHEL EMILY MARTIN/Examiner, Art Unit 1657