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
This application is a CON of 17/207,943 filed 03/22/2021 which is a CON of 16/322,956 filed 02/03/2019 PAT 10,982,252 which is a 371 of PCT/IL2017/050876 filed 08/08/2017 which claims benefit of 62/371,881 filed 08/08/2016. Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged.
Information Disclosure Statement
The information disclosure statement filed 12/19/2024 fails to comply with 37 CFR 1.98(a)(2), which requires a legible copy of each cited foreign patent document; each non-patent literature publication or that portion which caused it to be listed; and all other information or that portion which caused it to be listed. It has been placed in the application file, but the information referred to therein has not been considered.
Status of the Claims
Claims 1-20 are pending (claims set filed 09/20/2024) and are examined on the merits herein.
Claim Objections
Claim 7 is objected to because of the following informalities:
Claims 7 recites: “the nucleotide that encodes the enzyme” in line 3 and “the nucleotide that encodes the ubiquitinating enzyme target substrate” in line 6. Since nucleotide cannot encode an enzyme or an enzyme target substrate, Applicant is suggested to replace recitation of “nucleotide” with “polynucleotide”.
Appropriate correction is required.
Claim Rejections - 35 USC § 112
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 7-12 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 7 recites: “the 3’ end of the polynucleotide that encodes the first fragment of the second antibiotic resistance protein is linked to the 5’ end of the nucleotide that encodes the enzyme that is part of the ubiquitin pathway”. Claim 9 recites: ““the 3’ end of the polynucleotide encoding the N-terminal fragment of the DHFR protein is linked to the 5’ end of the polynucleotide capable of expressing the at least one enzyme that is part of the ubiquitin pathway”. The ubiquitinating enzymes are defined in the specification as referring to “ubiquitin-activating enzymes (E1s), ubiquitin-conjugating enzymes (E2s) or ubiquitin ligases (E3s)” (paragraph 0145). Although ubiquitin protein is a part of the ubiquitin pathway, ubiquitin is not an enzyme. However, claim 13 recites: “the at least one enzyme that is part of the ubiquitin pathway includes an E1 protein, an E2 protein, an E3 protein and a ubiquitin protein. Additionally, the specification does not describe E1, E2 or E3 enzymes to be fused to the second antibiotic resistance protein or DHFR but does describe ubiquitin attached to the reporter polypeptide fragment (paragraphs 0018, 0034, Figure 1A). It is not clear whether ubiquitin is included as part of the ubiquitin pathway in the limitations of claims 7 and 9. Therefore, the scope and boundaries of claims 7 and 9 are not certain making claims 7 and 9 indefinite.
Claim 8 dependent on claim 7 and claims 9-12 dependent on claim 8, do not resolve the issue mentioned above and are rejected.
For examination claims 7 and 9 are interpreted to include ubiquitin as the protein attached to the reporter fragment based on the specification and claim 13.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
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.
Claims 1, 2, 5-17 and 19 are rejected under 35 U.S.C. 103 as being unpatentable over Keren-Kaplan (Keren-Kaplan et al. The EMBO Journal, 2012, 31, 378-390) in view of Kerppola (US 7588907 B2) and Pelletier (Pelletier et al. PNAS, 1998, 95, 12141-12146).
Regarding claim 1, Keren-Kaplan teaches reconstruction of the entire functionally active eukaryotic ubiquitination cascade in bacteria: “… we reconstructed the entire eukaryotic ubiquitylation system in Escherichia coli.” (p. 378, right column, 3rd paragraph) and “…we have reconstituted in bacteria a pathway that specifically and accurately generates ubiquitylated eukaryotic proteins” (p. 387, left column last paragraph). The expressed system includes ubiquitin, E1-activating enzyme, E2-conjugating enzyme, E3 ligase and ubiquitylation substrate (p. 379, left column, 2nd paragraph). The system is composed of two expression vectors. The first vector has CDF origin of replication, pT7 promoter, polynucleotides encoding E1 protein, E2 protein and ubiquitin and a kanamycin antibiotic resistant protein. The second vector has ColE1 origin of replication, pTac promoter, polynucleotides encoding E3 protein and a protein – substrate of ubiquitylation and an ampicillin antibiotic resistant protein (p. 379, left column, 2nd paragraph, Figure 1). Keren-Kaplan discloses that the main advantage of expression of the ubiquitination enzymes in bacteria is the absence of deubiquitylation and mentions that it can be used for identification of new ubiquitylation substrates and to study ubiquitylation enzymes (p. 386, right column, 1st paragraph). Keren-Kaplan describes ubiquitylation of several proteins using the developed system (Abstract).
Keren-Kaplan does not teach vectors to include polynucleotides encoding fragments of another antibiotic resistant protein.
Kerppola teaches a fluorescent complementation assay for the detection of ubiquitinated proteins (Abstract). Kerppola states that the method can be used for screening of drugs affecting ubiquitination or ubiquitinated proteins, substrates of ubiquitination or in research: “The methods and compositions of the present invention find use in drug screening (e.g., to screen for drugs that alter ubiquitination or other modifications of proteins) and research (e.g., to identify ubiquitinated proteins and ubiquitination pathways).” (column 1, lines 48-52). Kerppola discloses that the invention method is based on complementation of split fluorescent protein, first fragment of which is fused to the first protein (ubiquitin) and the second to the second protein which can be protein being ubiquitinated, i.e. the ubiquitinating enzyme target substrate: “…a method, comprising providing a first fusion protein comprising a first fragment of a fluorescent protein linked to a first protein (e.g., ubiquitin) known to covalently modify a protein of interest; and a second fusion protein comprising a second fragment of the fluorescent protein linked to a protein of interest…” (column 2, lines 20-25) and “… the protein of interest is a protein suspected of being ubiquitinated…” (column 1, line 67, column 2, line 1). Upon covalent modification (ubiquitination) fragments of fluorescent protein associate and produce detectable signal: “… the first and second fragments of the fluorescent protein associate to form a fluorescent protein complex that generates an optically detectable signal in the presence but not the absence of covalent modification of the protein of interest by the first protein.” (column 2, lines 27-31). Kerppola mentions that the ubiquitin complementation assay can be performed in different host cells, including bacterial cells and comprising E. coli (column 15, lines 46-49). Thus, Kerppola teaches method for detection of ubiquitination using split peptide the first fragment of which is attached to ubiquitin and the second to ubiquitination substrate.
Pelletier teaches split antibiotic-resistance complementation assay based on murine dihydrofolate reductase (mDHFR). Pelletier discloses that the active mDHFR can be reassembled from complementary N- and C-terminal fragments when fused to either homodimerizing GCN4 leucine zipper-forming sequences or to heterodimerizing protein partners (Abstract). The assembly of mDHFR provides resistance to antibiotic trimethoprim because bacterial DHFR is inhibited by trimethoprim, while mammalian is not due to low affinity (p. 12143, right column, 4th paragraph). Therefore, determination of the DHFR reassembly and hence binding of protein partners can be detected in vivo by bacterial survival. Pelletier provides examples of detection of the interaction of protein partners, i.e. p21 ras-GTPase with its target RBD and immunophillin FKBP with TOR2 mediated by rapamycin, by bacterial cells survival assay (p. 12145, right column, 2nd paragraph). Pelletier mentions that the DHFR complementation assay can be used for studying protein-protein interaction since interaction of proteins is required for reconstitution of DHFR activity (p. 12146, left column, last paragraph).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify Karren-Kaplan expression vectors by introducing first fragment of fluorescent protein attached to ubiquitin into the first vector and the second fragment of the fluorescent protein attached to the ubiquitination substrate in to the second vector as taught by Kerppola. One would have been motivated to make this modification since split complementation assay described by Kerppola provides efficient detection of protein ubiquitination by detection of optical signal. A skilled artisan would have reasonably expected success in this combination since both Keren- Kaplan and Kerppola teach ubiquitination of proteins.
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify expression vectors designed based on Keren- Kaplan and Kerppola teachings for detection of ubiquitinating enzymes and ubiquitinated substrates by measurement of ubiquitination by complementation assay with the split fluorescent protein and use the split DHFR complementation assay described by Pelletier, i.e. substitute fragments of the fluorescent protein with fragments of DHFR. One would have been motivated to make this modification to facilitate detection of ubiquitination of protein test substrates by simple and fast assay of bacterial cell survival. A skilled artisan would have reasonably expected success in this modification since Kerppola and Pelletier describe application of split complementation assays based on the same principle but with different split proteins and hence detection method. Therefore, Keren-Kaplan provided expression vectors for complete ubiquitinating system expressed in bacteria, Kerppola provided complementation assay for detection of ubiquitinating enzymes and ubiquitinated substrates and Pelletier teaches faster and more simple survival assay for protein-protein interactions applicable to ubiquitination.
Thus, Keren-Kaplan, Kerppola and Pelletier teachings render claim 1 obvious.
Regarding claim 2, Keren-Kaplan teaches expression of expression vectors in bacteria (Abstract). Thus, Keren-Kaplan, Kerppola and Pelletier teachings render claim 2 obvious.
Regarding claims 5, and 6, Pelletier teaches DHFR as the split antibiotic resistance protein providing resistance to trimethoprim (p. 12143, right column, 4th paragraph).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to follow Pelletier teaching and use DHFR providing resistance to trimethoprim in complementation assay to detect ubiquitination based on Keren- Kaplan and Kerppola teachings. One would have been motivated to do so since DHFR provides resistance to trimethoprim antibiotic and ubiquitination of protein test substrates can be determined by cells survival. A skilled artisan would have reasonably expected success in that since Keren-Kaplan and Kerppola teach ubiquitination of proteins and Kerppola and Pelletier describe split complementation assays to detect protein-protein interactions. Thus, Keren-Kaplan, Kerppola and Pelletier teachings render claims 5 and 6 obvious.
Regarding claim 7, Kerppola teaches that the fragments of the fluorescent protein are fused to the 5’ end of ubiquitin and 5’ end of the ubiquitination substrate (column 22, lines 38-42) and suggests to fuse the second fluorescent fragment to different ends of the ubiquitination substrate (p. 25, lines 34-36). Regarding ubiquitin, please see interpretation of claim 7 above for 112b rejection. Pelletier teaches that one of the advantage of the assay is that the known structure of DHFR allows to control the protein-protein interaction by changing the orientation of interacting protein by fusing them to either N or C termini of DHFR (p. 12146, right column) and therefore also suggests different orientation for reporter protein fragments.
It would have obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to fuse the first fragment of DHFR from Pelletier teaching to the 5’-end of ubiquitin as taught by Kerppola for the first fragment of fluorescent reporter protein attached to ubiquitin in expression vector taught by Keren-Kaplan. One would have been motivated to do so with reasonable expected success because Kerppola and Pelletier teach split complementation assays for detection of protein-protein interactions, Kerppola teaches application of the complementation assay to ubiquitination and detection of protein-protein interactions in Pelletier teaching is based of DHFR reassembly providing cells survival that can be applied to detection of ubiquitination.
It would have obvious to one of ordinary skill in the art before the effective filing date of the claimed invention try different orientation of the second DHFR fragment fusion to ubiquitination substrate, i.e. either N-terminal or C-terminal. One would have been motivated to do so to select the optimal configuration since Kerppola and Pelletier suggest different configuration of fusion of complementation protein fragments with protein partners and Pelletier teaches that orientation of fusion protein can be changed based on known structure of DHFR. A skilled artisan would have reasonably expected success in that since and since optimization of the fusion protein orientation is within the skills of the artisan in the field. Thus, Keren-Kaplan, Kerppola and Pelletier teachings render claim 7 obvious.
Regarding claim 8, Pelletier teaches that fragment of DHFR were obtained by cutting the complete sequence of DHFR into N-terminal fragment and C-terminal fragment at residue 107 (p. 12143, right column, 3rd paragraph) and that fragments are fused to interacting protein partners (Abstract).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to follow Pelletier teaching and fuse N-terminal fragment to ubiquitin and C-terminal fragment to ubiquitination substrate. One would have been motivated to do so because ubiquitination of the substrate will result in DHFR reassembly that provides cell survival in the presence of trimethoprim and hence DHFR complementation assay facilitates detection of ubiquitination of protein test substrates by simple and fast assay. A skilled artisan would have reasonably expected success in that since Keren-Kaplan and Kerppola teach ubiquitination of proteins and Kerppola and Pelletier describe split complementation assays. Thus, Keren-Kaplan, Kerppola and Pelletier teachings render claim 8 obvious.
Regarding claims 9 and 10, Kerppola teaches that the fragment of fluorescent protein can be attached to the ubiquitin or substrate for ubiquitination via a linker sequence: “…each of the fusion proteins comprises a fragment of a fluorescent protein and a conjugation partner (e.g., ubiquitin or a candidate protein for ubiquitination), optionally joined by a linker sequence” (column 12, lines 61-64). Pelletier teaches that protein partners can be attached to DHFR reporter fragments via soluble linkers to allow optimal orientation for assembly (p. 12141, left column, last paragraph).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to follow Kerppola and Pelletier teaching and fuse fragments of the reporter protein to the interacting proteins via linkers, i.e provide linker between the first fragment of DHFR and the 5’ end of the ubiquitin and provide linker between the second fragment of DHFR and the ubiquitination substrate. One would have been motivated to do so because ubiquitination of the substrate will result in DHFR reassembly that provides cell survival in the presence of trimethoprim and Pelletier teaches that linkers will allow optimal orientation and assembly of DHFR. A skilled artisan would have reasonably expected success in that since Keren-Kaplan and Kerppola teach ubiquitination of proteins and Kerppola and Pelletier describe split complementation assays. Thus, Keren-Kaplan, Kerppola and Pelletier teachings render claims 9 and 10 obvious.
Regarding claims 11 and 12, Pelletier teaches that fragment of DHFR were obtained by cutting the complete sequence of DHFR into N-terminal fragment and C-terminal fragment at residue 107 which is in the disordered loop and can be disrupted with no significant effect on activity. Pelletier mentions that this fragmentation causes minimal disturbance to the active site (p. 12143, right column, 3rd paragraph).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to follow Pelletier teaching and use DHFR providing resistance to trimethoprim in complementation assay to detect ubiquitination based on Keren- Kaplan and Kerppola teachings and prepare fragments of complete DHFR protein cut at position not disturbing the active site and fuse N-terminal fragment to ubiquitin and C-terminal fragment to ubiquitination substrate. One would have been motivated to do so because ubiquitination of the substrate will result in DHFR reassembly that provides cell survival in the presence of trimethoprim and hence DHFR complementation assay facilitates detection of ubiquitination of protein test substrates by simple and fast assay. A skilled artisan would have reasonably expected success in that since Keren-Kaplan and Kerppola teach ubiquitination of proteins and Kerppola and Pelletier describe split complementation assays. Thus, Keren-Kaplan, Kerppola and Pelletier teachings render claims 11 and 12 obvious.
Regarding claims 13 and 14, Keren-Kaplan teaches the expressed system including ubiquitin, E1-activating enzyme, E2-conjugating enzyme, E3 ligase and ubiquitylation substrate (p. 379, left column, 2nd paragraph, Figure 1). Thus, Keren-Kaplan, Kerppola and Pelletier teachings render claims 13 and 14 obvious.
Regarding claims 15 and 16, Keren-Kaplan teaches expression of human ubiquitin (p. 387, right column, 2nd paragraph). Thus, Keren-Kaplan, Kerppola and Pelletier teachings render claims 15 and 16 obvious.
Regarding claim 17, Keren-Kaplan teaches the first expression vector to have CDF as origin of replication and the second expression vector – ColE1 as origin of replication. Thus, Keren-Kaplan, Kerppola and Pelletier teachings render claim 17 obvious.
Regarding claim 19, Keren-Kaplan teaches that malfunctions in the ubiquitination system “underlie various human disorders including cancer and neurodegenerative, metabolic and infectious diseases (p. 378, right column, 1st paragraph). Keren-Kaplan discloses that the expression system can be used to study ubiquitination of human disease proteins including identification of their modification (p. 378, right column, last paragraph) indicating identification of the level of ubiquitinating enzyme target substrate ubiquitination. Thus, Keren-Kaplan, Kerppola and Pelletier teachings render claim 19 obvious.
Claim 3 is rejected under 35 U.S.C. 103 as being unpatentable over Keren-Kaplan (Keren-Kaplan et al. The EMBO Journal, 2012, 31, 378-390) in view of Kerppola (US 7588907 B2) and Pelletier (Pelletier et al. PNAS, 1998, 95, 12141-12146) as applied to claim 1 above, and further in view of Tuvia (US 20060188890 A1).
The teachings of Keren-Kaplan, Kerppola and Pelletier have been set forth above.
The teachings of Keren-Kaplan, Kerppola and Pelletier do not teach a third expression vector.
Tuvia teaches identification of substrates for ubiquitination employing ubiquitin-containing fusion protein and substrate-containing fusion protein. Each fusion protein expressed from a separate expression vector contains output-generating domain such that upon bond formation between ubiquitin and the substrate, the two domains are brought into close proximity and the output signal is generated (paragraph 0008, 0019). Tuvia discloses that the method can include the step to determine whether output signal/ubiquitination depends on the presence of E3 (paragraph 0010). Tuvia describes that the nucleic acid encoding E3 can be on the expression vector comprising inducible promoter operably linked to E3 coding sequence (paragraph 0015). Tuvia mentions that nucleic acid encoding E3 can be part of the first expression vector or E3 can be expressed from a third expression vector (paragraphs 0020, 0021).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify 2 expression vector expression system based on Keren- Kaplan, Kerppola and Pelletier teachings and express E3 from a third expression vector as taught by Tuvia. One would have been motivated to do so because that would allow to control expression of E3 by inducible promoter separately from expression of other proteins of ubiquitination system to determine whether ubiquitination depends on E3. A skilled artisan would have reasonably expected success in that since Keren-Kaplan, Kerppola and Tuvia teach ubiquitination and Kerppola, Pelletier and Tuvia describe fusion proteins to detect protein interaction. Thus, Keren-Kaplan, Kerppola, Pelletier and Tuvia teachings render claim 3 obvious.
Claims 4, 18 and 20 are rejected under 35 U.S.C. 103 as being unpatentable over Keren-Kaplan (Keren-Kaplan et al. The EMBO Journal, 2012, 31, 378-390) in view of Kerppola (US 7588907 B2) and Pelletier (Pelletier et al. PNAS, 1998, 95, 12141-12146) as applied to claims 1, 5-14, 16 and 17 above and further in view of Peteranderl (Peteranderl et al. Appl. Envir. Microbiol., 1990, 56, 1981-1983).
The teachings of Keren-Kaplan, Kerppola and Pelletier have been set forth above.
As described above for claims 1, 5-14, 16 and 17, Keren-Kaplan teaches the first vector comprising CDF origin of replication, polynucleotides encoding E1 protein, E2 protein and ubiquitin and a kanamycin antibiotic resistant protein and the second vector comprising ColE1 origin of replication, polynucleotides encoding E3 protein and a protein – substrate of ubiquitylation and an ampicillin antibiotic resistant protein (p. 379, left column, 2nd paragraph). Kerppola teaches fragments of reporter protein fused to ubiquitin of the first vector and to the ubiquitination substrate on the second vector (column 1, line 67, column 2, lines 1, 20-25). Pelletier teaches DHFR as a reporter protein for the complementation assay fragments of which are fused to interacting proteins and reassembly of which provides cells survival and indicates interaction between fused proteins (p. 12143, right column, 4th paragraph). Both Kerppola (column 12, lines 61-64) and Pelletier (p. 12141, left column, last paragraph disclose presence of linkers between the reporter protein fragments and the interacting proteins. Kerppola describes attachment of the first reporter protein fragment to 5’end of ubiquitin and suggests to fuse the second fluorescent fragment to different ends of the ubiquitination substrate and Pelletier suggests changing the orientation of interacting protein by fusing them to either N or C termini of DHFR (p. 12146, right column) providing motivation to try different orientation. Thus, combination of prior art of Keren-Kaplan, Kerppola and Pelletier provides the claimed first and second vector except that Keren-Kaplan, Kerppola and Pelletier do not teach antibiotic resistance protein conferring resistance streptomycin on the second vector.
Regarding claim 4 and 18, Peteranderl teaches comparison of the stability of various antibiotics (Abstract). Table 1 shows degradation constants for antibiotic incubated in anaerobic medium and values are listed in order of decreasing stability (p. 1981, right column, 3rd paragraph, p. 1982, Table 1). It can be seen that ampicillin has the lowest stability in all conditions tested. Streptomycin, one of the commonly used antibiotics has higher stability than ampicillin.
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to replace the ampicillin resistant gene in the second expression vector based on combination of Keren- Kaplan, Kerppola and Pelletier teachings with another antibiotic resistant gene and try streptomycin based on Peteranderl teaching. One would have been motivated to do so because Peteranderl showed that stability of streptomycin is much higher than that of ampicillin and that would reduce amount of antibiotic used and provide more reliable results of ubiquitylation. A skilled artisan would have reasonably expected success in that since selection of antibiotic and replacement of antibiotic resistance gene is within the skills of the artisan in the field. Thus, Keren-Kaplan, Kerppola, Pelletier and Peteranderl teachings render claims 4 and 18 obvious.
Regarding claim 20, Kerppola teaches kits to perform ubiquitin mediated complementation assays containing first expression vector encoding first fusion protein of ubiquitin with the first reporter protein fragment and the second expression vector encoding the second fusion protein of ubiquitination substrate fused with the second reporter protein fragment. Kit can contain other components necessary for the assay including controls, directions, any necessary software for analysis and presentation of results (column 19, lines 66-67, column 20, lines 1-4, 6-9).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention that composition comprising two expression vector for ubiquitination detection based on Keren-Kaplan, Kerppola, Pelletier and Peteranderl teachings can be used as a kit for identification of ubiquitylation substrates and study ubiquitylation enzymes. One would have been motivated to assume so with the reasonably expected success because Kerppola describes kit including components for the complementation ubiquitylation assay, Keren-Kaplan provides expression vectors comprising complete ubiquitination system to be expressed in bacterium and Pelletier points to DHFR reporter for the complementation assay with detection by cell survival. Thus, Keren-Kaplan, Kerppola, Pelletier and Peteranderl teachings render claim 20 obvious.
Conclusion
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to LIOUBOV G KOROTCHKINA whose telephone number is (571)270-0911. The examiner can normally be reached Monday-Friday: 8:00-5:30.
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If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Sharmila G Landau can be reached at (571)272-0614. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/L.G.K./Examiner, Art Unit 1653
/SHARMILA G LANDAU/Supervisory Patent Examiner, Art Unit 1653
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