28 results about "Post translational" patented technology

This disclosure provides nanoparticle compositions, and use thereof for isolating and measuring proteins, protein degrader action, and cells. The disclosed nanoparticles can be identified by barcodes that reveal the identity of and post translational modifications to the bound protein using image processing techniques described herein.

The present invention relates to a rep plasmid comprising at least one long adeno-associated virus replication protein coding sequence encoding at least one long functional rep protein, at least one short adeno-associated virus replication protein coding sequence encoding at least one short functional rep protein, and at least one heterologous element, wherein following translation the at least one heterologous element results in a ratio of long adeno-associated virus rep protein to short adeno-associated virus rep protein of about 1:5 to about 1:5000.

The present disclosure relates to the use of a maltose dependent degron to control stability of a protein of interest fused thereto at the post-translational level. The present disclosure also relates to the use of a maltose dependent degron in combination with a maltose-responsive promoter to control gene expression at the transcriptional level and to control protein stability at the post-translational level. The present disclosure also relates to the use of a stabilization construct that couples expression of a cell-growth-affecting protein with the production of non-catabolic compounds. The present disclosure further relates to the use of a synthetic maltose-responsive promoter. The present disclosure further provides compositions and methods for using a maltose dependent degron, a maltose-responsive promoter, and a stabilization construct, either alone or in various combinations, for the production of non-catabolic compounds in genetically modified host cells.

The invention discloses a micropterus salmoides gene modification method which comprises the following steps: step 1, encoding a precursor gene of micropterus salmoides GLP according to a published micropterus salmoides genome database; 2, obtaining a translated micropterus salmoides GLP precursor protein sequence according to the precursor gene for coding the micropterus salmoides GLP; step 3, successfully predicting a mature protein sequence of the micropterus salmoides GLP according to a comparison result of the cloned sequence for coding the micropterus salmoides GLP precursor gene, a translated protein sequence, a recognition site of 1 / 3 of a prohormone converting enzyme and sequences of different fish GLPs; and 4, transforming the mature protein sequence of the GLP of the micropterus salmoides according to the mature protein sequence of the GLP of the micropterus salmoides and the recognition site of the DPPIV enzyme. The GLP is modified according to a mature protein sequence of the GLP, and the modified protein can realize a regulation effect on fish glycolipid metabolism and is expected to be developed into a feed additive for promoting glycolipid utilization.

An engineered tyrosyl-tRNA synthetase / tRNA pair that co-translationally incorporates O-sulfotyrosine in response to UAG codons in E. coli and mammalian cells is described herein. This platform enables recombinant expression of eukaryotic proteins homogeneously sulfated at chosen sites.

Proteases of Group IV (+)ssRNA viruses were found to act on a human sequences in addition to the viral sequences. The identity of the cleavable human sequences is disclosed. Detection of these sequences can act as a diagnostic of infection. It is contemplated that these findings could be employed to facilitate post-translational silencing at the level of protein (e.g., removal of existing proteins), thus serving as a protein analog to CRISPR / Cas9 and RNAi / RISC, and further to enable sequence-specific silencing of host functions without the modification of the host genome.

The application discloses a tobacco nicotine metabolism and plant height regulation related gene NtSULTR3.4 in nicotine metabolism and plant height regulation, and the CDS sequence of the NtSULTR3.4 is shown as SEQ ID No. 1. After the sequence of the NtSULTR3.4 gene is translated, the amino acid sequence of the encoded protein is shown as SEQ ID No. 2. By using a CRISPR / Cas9 mediated gene editing technology to knock the NtSULTR3.4 gene, edited materials with increased nicotine content and changed plant height are obtained, indicating that the gene is a negative regulatory factor of nicotine metabolism, and is positively correlated with the tobacco plant height, which provides genetic materials and theoretical basis for directional improvement of a new tobacco variety of tobacco nicotine metabolism gene function research, nicotine content regulation and plant height regulation.

The present disclosure relates to the use of a maltose dependent degron to control stability of a protein of interest fused thereto at the post-translational level. The present disclosure also relates to the use of a maltose dependent degron in combination with a maltose-responsive promoter to control gene expression at the transcriptional level and to control protein stability at the post-translational level. The present disclosure also relates to the use of a stabilization construct that couples expression of a cell-growth-affecting protein with the production of non-catabolic compounds. The present disclosure further relates to the use of a synthetic maltose-responsive promoter. The present disclosure further provides compositions and methods for using a maltose dependent degron, a maltose-responsive promoter, and a stabilization construct, either alone or in various combinations, for the production of non-catabolic compounds in genetically modified host cells.

The present invention relates to novel long non-coding RNAs (lncRNA) involved in the regulation of hypertrophy of chondrocytes, wherein RUNX2 was regulated at the post-translational level during hypertrophic differentiation of the chondrocyte cell line TC28a2 and two novel RUNX2-binding lncRNAs among numerous lncRNAs were identified, leading to the present invention. It is expected that bone-related diseases can be diagnosed by measuring the expression of the identified specific lncRNAs, and furthermore, the vascularization or calcification of articular cartilage can be effectively suppressed by inhibiting the hypertrophy of chondrocytes through inhibiting the expression of the identified specific lncRNAs, which is expected to be highly effective in the prevention or treatment of bone-related diseases.

The invention relates to a saccharomyces cerevisiae strain for producing 2, 3-butanediol through bidirectional light regulation and control as well as a construction method and application of the saccharomyces cerevisiae strain. The Saccharomyces cerevisiae strain for producing 2, 3-butanediol through bidirectional light regulation and control is named as BYSDBSPP, contains a PSD and SULI double-light-control system, and respectively regulates and controls an ethanol synthesis route and a 2, 3-butanediol synthesis route. The method is used for producing 2, 3-butanediol in a lysine-deficient SD liquid culture medium under the condition of light stimulation; the wavelength range of the light stimulation is 400 to 500 nm. According to the invention, the problem of bacterial strain growth defects caused by gene knockout is avoided, and the production of various target metabolites taking ethanol as a byproduct is facilitated. The light regulation system for controlling the protein level is adopted, the stability of specific zymoprotein in a metabolic pathway can be directly controlled from the post-translational level, and in metabolic engineering application, compared with a light regulation system for controlling the gene transcription level, the response speed is higher, and strain modification application is simpler and more convenient.

The present disclosure is directed to systems and methods for synthesizing a spidroin. In some embodiments, the methods comprise synthesizing a monomer in vivo in a heterologous host, the monomer comprising an N-terminus IntC domain and a C-terminus IntN domain, and post-translationally polymerizing the synthesized monomer via in vitro split-intein mediated polymerization.

The present application relates to mutated RNA polymerases (RNAP) enzymes from bacteriophage T7 that are post-translationally controlled by one or more to heterocyclic ligands, such as indole, an indole derivative, or an indole-similar compound. A preferred mutant includes a T7 RNAP with a mutation at position 727, and more preferably a thermostable T7 RNAP with a tryptophan to glycine substitution at position 727 of the amino acid sequence.

The invention relates to methods of controlling post-translational modification of a recombinant protein In brief, the methods comprising contacting a recombinant protein and / or a cell capable of producing the recombinant protein with one or more sulfinyl group-containing compounds, thereby controlling post-translational modification. The invention further relates to purified antibodies obtained by the methods as described herein.

Compounds, compositions, and methods are provided for covalently linking an antibody or an antibody fragment to a cargo molecule, such as a therapeutic or a diagnostic agent, using a combination of enzymatic glycan remodeling and click chemistry. The method allows a cargo molecule to be selectively and efficiently attached post-translationally to an antibody or an antibody fragment. Also provided are antibody drug conjugates, methods of making, and uses thereof.

Methods are provided for detecting disease in liquid biopsy samples by identifying microbial rRNA. The methods include isolating nucleic acids from extracellular vesicles, treating the nucleic acids with DNase to remove DNA including human cfDNA, mcfDNA, and contaminating microbial DNA, and then amplifying a region of a microbial phylogenetic rRNA. To further reduce contaminating DNA, the amplifying step can include reverse transcription to translate the phylogenetic region into cDNA and extend the cDNA ends with a T7 promoter and sequencing adaptors. The translated region is amplified with primers to the extended ends of the cDNA and sequenced. The resulting phylogenetic microbial sequences are useful for diagnosis and monitoring of diseases including cancer. The method can further include amplification of other human or bacterial DNA or RNA sequences to augment disease diagnosis. The method results in a very low contribution of contaminating microbial DNA and RNA to the overall microbial signature.

The invention provides a genetic expression cassette comprising, in order from 5'-3':an 5' adenoassociated viral (AAV) inverted terminal repeat (ITR), the elongation factor 1 long promoter, a chimeric intron, a 5' UT translational enhancer element containing a Kozak sequence, a polynucleotide comprising a nucleic acid sequence with at least 80% identity to the nucleic acid sequence of either wild-type of propionyl-CoA carboxylase beta (PCCB or synPCCB1), the hepatitis B virus derived post-translational response element, the bovine growth hormone poly-adenylation sequence, and an 3' AAV inverted ITR. The invention also provides expression vectors comprising the inventive cassette, compositions comprising the same, and methods for treating a disease or condition mediated by propionyl-CoA carboxylase.

A synthetic DNA molecule comprising one segment encoding a tumor neoantigen or an epitope from an infectious agent under the control of a promoter for the transcription into a corresponding RNA molecule and a segment for the translation of the translated RNA molecule into a peptide.

The invention provides an application of a signal peptide and / or an endoplasmic reticulum / cytoplasmic transmembrane region (ECTD) in preparation of a vaccine, and also provides an expression cassette and a polypeptide translated by the expression cassette, and the expression cassette encodes the signal peptide and / or the endoplasmic reticulum / cytoplasmic transmembrane region (ECTD) and a nucleic acid sequence of an antigen peptide. The invention also provides a vaccine containing a nucleic acid sequence for coding the expression cassette or a polypeptide obtained by translating the expression cassette and application of the vaccine in preparation of medicines for preventing or treating diseases.

This invention discloses a method and application for constructing transgenic silkworm lines that enhance the triple-helix structure of recombinant full-length human type III collagen using a binary hybridization strategy. First, this invention constructs a transgenic silkworm line co-expressing human prolyl 4-hydroxylase (hP4H) α and β subunits, and a transgenic silkworm line expressing recombinant full-length human type III collagen (rhCOLⅢ). Then, the two lines are genetically hybridized using a binary hybridization strategy, and double-positive offspring co-expressing rhCOLⅢ and hP4H are selected, achieving post-translational hydroxylation modification of rhCOLⅢ by hP4H. The modified recombinant full-length human type III collagen (rhCOLⅢ-Hyp) triple-helix structure content is significantly increased. This invention provides a key modification tool and technical pathway for producing full-length recombinant human collagen with a complete triple-helix structure using a silkworm bioreactor.

The invention discloses application of morusin O in preparation of a TEAD protein palmitoylation inhibitor, and relates to the technical field of molecular biology. The invention also provides a TEAD palmitoylation inhibitor and a method for screening the TEAD palmitoylation inhibitor in vitro. It is found for the first time that the compound can be directly combined with a lipid binding pocket of TEAD1 protein to interfere the post-translational palmitoylation modification level of the TEAD1 protein, so that the conformational stability and transcriptional activity of TEAD1 are reduced, and the application value is wide.

The present disclosure generally relates to multi-chain chimeric polypeptides or receptors having distinct polypeptide chains that associate post-translationally to enable the simultaneous activation of the signaling domain and the release of a transcriptional regulator upon binding of a ligand. The disclosure also provides nucleic acid constructs, recombinant cells, vectors, pharmaceutical compositions, and methods of treatment including the multi-chain chimeric polypeptides of the disclosure. The disclosure further provides methods for simultaneously inducing T cell signaling and gene regulation in a T cell.

The invention discloses application of an FMR1 inhibitor in preparation of a medicine for treating immunological rejection type gastric cancer. Experimental studies prove that FTO is continuously and highly expressed in immunological rejection phenotypes and is related to interstitial activation and T cell rejection. Continuous immunohistochemistry and multiple immunofluorescence show that the FMR1 protein is highly expressed in an immunological rejection type tumor microenvironment, and most CD8 + T cells are located in tumor interstitial substances. In mechanism, the FMR1 regulates and stabilizes FTO protein and limits the degradation of proteasome of the FTO protein through post-translation, so that FTO-dependent m6A reprogramming is maintained, and an immunological rejection microenvironment is enhanced. Therefore, the invention discloses a non-classical mechanism that the FMR1 modifies and stabilizes the FTO protein after translation, and the FMR1-FTO axis can be used as a potential intervention target for breaking an immune barrier and sensitizing gastric cancer immunotherapy.

The invention discloses a recombinant bovine leukemia virus infectious clone plasmid, a preparation method and application, the recombinant plasmid is based on the bovine leukemia virus infectious clone plasmid, and an IRES element and a target gene are inserted into a non-coding region between an Env gene and a Tax gene of a bovine leukemia virus fragment. When the recombinant bovine leukemia virus infectious clone plasmid is used for transfecting cells and then culturing, the recombinant bovine leukemia virus capable of stably expressing a target gene can be successfully rescued and obtained, the protein processing modification after the infectious clone plasmid translation is not influenced, and the assembly of filial generation virus particles is not influenced; the expressed progeny virus can efficiently express a target gene, can be stably inherited in continuous passage, and has a relatively high application value.

The present application relates to polypeptides, in particular to an animal toxin polypeptide and application thereof. The polypeptide is identified by RNA sequencing of centipede transcriptome, and bioinformatics screening and Motif scanning analysis of translated protein sequence. Experimental results show that SM-8 shows strong analgesic effect in various pain models, including hot plate test, acetic acid writhing test, formalin inflammatory pain model and migraine model, and the analgesic effect is equivalent to that of butorphanol tartrate and sumatriptan. Molecular mechanism research shows that SM-8 can regulate the expression of pain-related genes. Preliminary safety evaluation by mouse open field test shows that the polypeptide does not cause obvious toxic side effects, has good safety, and has good prospects for development into a new polypeptide analgesic drug.

The invention provides a protein labeling method and application thereof, and belongs to the technical field of biology. The method comprises the steps that N mRNA sequences are obtained, the 3'ends of the mRNA sequences are provided with marker sequence binding sites, and the marker sequences comprise a fixed sequence and at least one bar code sequence; the fixed sequence has a first modification and a second modification; the first modification is used for covalently linking the marker sequence with the protein translated by the mRNA sequence; the second modification is used for fixing the marker sequence on the mRNA sequence; the bar code sequences of the ith mRNA sequence and the jth mRNA sequence are different; n is an integer not less than 1, and i and j are integers not less than 1 and not greater than N; hybridizing the fixed sequence, the bar code sequence and the N mRNA sequences to obtain N marker compounds; and translating the N labeled complexes to obtain N labeled proteins. The method can realize simultaneous labeling of batches of proteins.

The present disclosure provides programmable base editors, and methods for their production and use. A subject programmable base editor includes (1) a CRISPR-Cas fusion protein comprising a CRISPR-Cas effector protein fused to a first member of a split-protein binding pair, where the CRISPR-Cas effector protein is a nickase (e.g., nCas9) or is catalytically inactive (e.g., dCas9, dCas12 such as dCas12a), and (2) a deaminase fusion protein comprising a deaminase protein fused to a second member of the split-protein binding pair. The CRISPR-Cas effector protein and the deaminase protein are not translationally fused to one another, but are instead post-translationally bound, e.g., in some cases covalently linked via a covalent bond (e.g., an isopeptide bond) formed between the two members of the split-protein binding pair (e.g. between a catcher protein and a partner tag protein).

This invention relates to providing technical tools, such as nucleic acids, which, after transcription or translation in plants, are suitable for mediating the characteristics of haploid inducible lines or for increasing the inducibility of haploid inducible lines, as well as methods and uses for generating and identifying non-transgenic and transgenic plant haploid inducible lines, and to improving existing plant haploid inducible lines.

The invention provides a MAFG transcriptional activation luciferase reporting system based on an MYC promoter, relates to the technical field of biomedicine, and is technically characterized in that the MAFG transcriptional activation luciferase reporting system based on the MYC promoter is provided and comprises a reporter gene plasmid constructed by an MYC promoter truncation body and firefly luciferase, the MYC promoter truncation body comprises a sequence as shown in SEQ ID NO: 01. The invention provides a dual-luciferase report system (Dual-Luciferase report) based on an MYC promoter, and the dual-Luciferase report system (Dual-Luciferase report) is used for detecting the activation transcription capability of a transcription factor MAFG. The luciferase does not need post-translation processing, so that report activity is generated immediately once translation is carried out; and the light product quantum efficiency is high, so that the detection is sensitive and rapid. The method is mainly applied to miRNA target gene verification, transcription factor regulation verification and the like at present. The system can accurately reflect the transcriptional activity of MAFG based on luciferase of an MYC promoter region.