2848 results about "Protein expression" patented technology

The invention provides a multielement polyA polynucleotide for improving mRNA (messenger ribonucleic acid) stability and protein expression and application of the multielement polyA polynucleotide. The invention firstly provides a polynucleotide, the polynucleotide comprises more than two sections of polyA sequence fragments, the adjacent polyA sequence fragments are connected through Linkers, each polyA sequence fragment is composed of one or more continuous A, and each Linker is composed of C and / or G. The polynucleotide provided by the invention can improve mRNA stability and protein expression, and compared with a traditional unit poly-A sequence, the polynucleotide provided by the invention can greatly reduce the recombination rate of fermentation in escherichia coli so as to more easily optimize and amplify the process, and can be better applied to plasmid fermentation related to transcription template preparation in mRNA vaccine or drug production.

The invention provides a protein expression prediction method based on a convolutional neural network and a residual attention mechanism, and belongs to the field of bioinformatics and artificial intelligence. The method comprises the following steps: performing data cleaning and binary one-hot coding on a DNA sequence, inputting the coded DNA sequence into a feature extraction layer, and extracting local features of DNA; local features are extracted through a convolutional neural network, global features are obtained through a multi-head attention mechanism, and residual connection is combined; and finally, inputting the global features into a full connection layer to generate a protein expression prediction result. Furthermore, an SHAP interpretability method is utilized to interpret the output result of the prediction model, and the contribution of the basic group at each position in the DNA sequence to the predicted value is quantified. According to the method, deep learning and interpretability technologies are combined, the precision and credibility of protein expression prediction are remarkably improved, and the method is suitable for predicting protein expression.

The invention discloses a drug resistance prediction method and system based on comparative learning and multi-modal fusion, and the method comprises the steps: firstly generating a molecular map and a molecular fingerprint based on the SMILES of a target drug, and extracting the molecular features of the drug through a comparative learning model constructed through combining a map attention network and a map convolution network; and then, acquiring protein expression, gene expression and metabolic expression data from the target tissue cells, extracting modal features through a deep convolutional network, a Transform encoder and a multi-dimensional attention network, and realizing adaptive fusion of the multi-modal features through a heterogeneous interactive attention mechanism. And finally, jointly inputting the fused multi-modal features and drug molecular features into a multi-layer sensor to realize high-precision prediction of the drug resistance of cells to drugs. By introducing a contrast learning and multi-modal feature fusion mechanism, the characterization capability and prediction precision of the model are effectively improved, and efficient and reliable support can be provided for drug screening and clinical decision making.

The invention belongs to the field of biological medicines, and particularly relates to a varicella-zoster virus gE fusion protein as well as a preparation method and application thereof. The varicella-zoster virus gE fusion protein comprises an amino acid sequence as shown in SEQ ID NO: 1 and at least one of the following fragments: a signal peptide, a Foldon sequence, a Ferritin sequence and a His tag sequence, the signal peptide can improve the protein expression quantity, the Foldon sequence can improve the stability of the protein, the particle size of an antigen can be increased, the immunogenicity of the antigen can be improved, and the fusion protein can be used for preparing a medicine for treating the varicella-zoster virus gE fusion protein. The Ferritin sequence can enhance immune response, a vaccine prepared from the gE fusion protein has excellent immunogenicity, and compared with a commercially available vaccine Shingrix, higher humoral immunity and cellular immunity levels can be induced.

The invention belongs to the technical field of molecular biology and genetics, and particularly relates to a method for editing cattle genes based on Pro-iCHI. The invention provides a method for editing cattle genes based on Pro-iCHI, which comprises the following steps: transiently expressing protamine in cattle AG-haESCs after gene editing, and injecting into mature oocytes to obtain reconstructed embryos. The protamine can remove abnormal DNA methylation generated by an oocyte cytoplasm injection haploid stem cell technology, cell nucleuses are compressed into a sperm-like structure, the obtained bovine Pro-iCHI embryo can smoothly develop into a blastocyst, and the ratio of the blastocyst to the blastocyst obtained by in-vitro fertilization is equivalent. Moreover, the protamine coding gene is inserted into the saccharomyces cerevisiae protein expression vector and then is subjected to transient expression, so that the protamine coding gene cannot be integrated into a genome to cause insertion of an exogenous gene while abnormal DNA methylation is erased.

The invention relates to the technical field of protein prediction, and discloses a protein interaction prediction method based on cross-modal enhanced representation learning. According to the method, a Convform module and a Graphform module are designed, and a global attention mechanism is introduced into each module, so that the limitation that only local features or global features are concerned in an existing method is broken through. According to the design, the model can capture the local mode and the global dependency relationship of the protein at the same time, and more comprehensive protein representation is provided. The invention provides a cross-modal enhancement method based on comparative learning. The problem of difference between a sequence mode and a structural mode is solved through an explicit mode alignment process. The method not only can maintain unique information of each mode, but also can give full play to complementarity among the modes, so that richer protein expression is obtained. The cross-protein fusion module provided by the invention can more accurately capture essential characteristics of protein interaction, and the accuracy of PPI prediction is remarkably improved.

The invention provides an UTR (Untranslated Region) element H2202 P1-G as well as a construction method and application thereof, and relates to the technical field of mRNA (messenger ribonucleic acid). According to the present invention, the ribosome load prediction and the secondary structure optimization are performed on the natural 5 'UTR of the HIV TAT 202 gene through the BaidleHelix platform, and the obtained HTAT 202 P1 sequence avoids the inhibitory hairpin structure so as to significantly improve the luciferase expression quantity compared to the natural UTR; an ncRNA sequence without a secondary structure is introduced on the basis of the HTAT 202 P1, translation inhibition of a 5 'cap region is further relieved, and the protein expression quantity of the constructed H2202 P1-G mutant (the DNA sequence of the H2202 P1-G is as shown in SEQ NO 1, and the RNA sequence is as shown in SEQ NO 2) is further improved.

The invention discloses a method for analyzing the tumor microenvironment of esophageal squamous cell carcinoma. The method comprises the following steps: selecting a basic tissue structure protein to be analyzed in a tumor microenvironment, carrying out cell segmentation and protein expression level extraction, carrying out clustering identification on cell types, identifying a three-level lymph structure and a neighborhood thereof, and determining an immune cell proportion and an expression level of a key functional protein. Identifying tissue regions and neighborhoods thereof and determining different cell proportions and expression levels of key functional proteins, as well as determining cell-cell spatial interactions in the tumor microenvironment. The method provided by the invention can greatly improve the efficiency and accuracy of predicting the intervention effect.

The invention relates to the technical field of recombinant protein expression, in particular to recombinant human-derived III-type collagen as well as a preparation method and application thereof. The invention provides a recombinant human source III type collagen. The amino acid sequence of the recombinant human source III type collagen is shown as SEQ ID NO. 1. The recombinant human-derived III-type collagen prepared by the invention can be better applied to a human body, better exerts biological functions of the collagen, avoids repulsive interaction, is high in safety, can effectively promote cell proliferation and oxidation resistance, and is high in expression yield, stable in batches and good in quality; the recombinant human III type collagen prepared by the invention has the potential of being applied to preparation and production of biological medicines and cosmetics.

The invention discloses application of an RNF130 activator in preparation of a medicine for preventing and / or treating ischemic heart disease, and belongs to the technical field of biological medicine. Research results show that when an RNF130 activator is given in a mouse of a myocardial ischemia reperfusion injury model, the left ventricular ejection fraction and the left ventricular short-axis shortening rate of the mouse can be improved, myocardial tissue pathological injury can be relieved, the protein expression levels of NOX-2, BAX and TNF-alpha are reduced, the myocardial infarction area is reduced, and myocardial cell apoptosis is reduced; therefore, the RNF130 activator has prevention and treatment effects on the myocardial ischemia-reperfusion injury, that is, the RNF130 activator can be used as an effective component of the medicine for preventing or treating diabetes complicated by myocardial ischemia-reperfusion injury, and a new strategy is provided for preparation of the medicine for preventing and treating myocardial ischemia-reperfusion injury.

Relating to the technical field of gene therapy, provided is an siRNA molecule for inhibiting PCSK9 expression, and use thereof. Particularly, an siRNA molecule for inhibiting PCSK9 expression (comprising a sense strand and an anti-sense strand that are complementary to form a double-stranded region) and a pharmaceutical composition comprising the same are disclosed. The siRNA molecule has a high silencing efficiency and a long-lasting silencing effect for PCSK9 gene, and is useful in the manufacture of products for inhibiting PCSK9 protein expression, products for reducing the concentration of low-density lipoprotein or low-density lipoprotein cholesterol in serum; and products for relieving the symptoms of PCSK9 gene-mediated diseases.

The invention discloses application of an ALOX15B gene serving as a target spot to screening of medicines for treating pulmonary vascular remodeling and pulmonary arterial hypertension, and belongs to the technical field of biological medicines. The invention discloses application of an ALOX15B gene or protein expression inhibitor in preparation of a medicine for treating pulmonary vascular remodeling and pulmonary arterial hypertension. By inhibiting expression of the ALOX15B gene or the encoding protein thereof, pulmonary vascular remodeling can be effectively reversed, and rising of pulmonary arterial pressure caused by oxygen deficit can be effectively reversed, so that the ALOX15B gene can be applied to treatment of pulmonary arterial hypertension related diseases.

Disclosed herein are chimeric antisense compounds and methods for reducing HBV mRNA, DNA, and protein expression. Such methods, compounds, and compositions are useful for treating, preventing, or ameliorating HBV-related diseases, disorders, or conditions.

The invention relates to the technical field of biological fermentation, in particular to a methanol-induced fermentation process for expressing III type recombinant human collagen, which comprises the following steps: inoculating a pichia pastoris fermentation seed solution into a fermentation culture medium, performing fermentation culture to a glycerol culture stage, and entering a methanol-induced culture stage after glycerol feeding is finished and pH is rebounded. According to the methanol-induced fermentation process for expressing the III-type recombinant human collagen, provided by the invention, after the glycerol feeding is finished, the pH rebound is taken as an index, and methanol induction is started after the pH rebound, so that thallus growth is obviously accelerated, and the protein expression quantity is improved, and therefore, the III-type recombinant human collagen can be industrially produced; and greater economic values can be created for enterprises.

The invention discloses a method for aided analysis of influence of 5 'UTR on mRNA translation efficiency based on a machine learning model, and relates to the technical field of synthetic biology and computational biology, the method comprises the following steps: S1, constructing a 5' UTR plasmid library; s2, transcribing the 5 'UTR plasmid into an independent mRNA (messenger ribonucleic acid) with a luciferase-opt luciferase, and taking the luminous intensity of the luciferase-opt luciferase as the translation intensity of the mRNA; s3, analyzing characteristics related to protein expression intensity of the mRNA; and S4, constructing a random forest algorithm model on the basis of the features analyzed in the step S3 to predict the influence of the 5 'UTR on the mRNA translation efficiency. According to the method for auxiliary analysis of the influence of the 5 'UTR on the mRNA translation efficiency based on the machine learning model, a feasible technical route reference is provided for sequence optimization of mRNA in subsequent mRNA vaccine development, so that the mRNA vaccine can efficiently express target protein in a body.

Provided herein are methods of cell-free protein synthesis, optimised cell-free protein synthesis (CFPS) reagents for the expression of eukaryotic proteins and methods for optimising CFPS reagents to increase protein expression yields for mammalian proteins and subsequent expression in cells. The methods are applicable to protein expression on a microfluidic device having hydrophobic surfaces by merging droplets on the device in order to screen a selection of expression compositions in parallel and then identify the optimum conditions for producing the stable folded protein in a cell-based system.

The present invention relates to a method for constructing an eight-gene-edited xenogeneic organ transplantation donor pig, belonging to the field of animal biotechnology. In the wild-type porcine fetal fibroblast cell line, the GGTA1, β4GalNT2, and CMAH genes are knocked out by using the CRISPR / Cas9 gene editing technology, and the humanized genes of hCD39, hCD46, hCD55, hCD59, and hTBM are transfected. Combining with somatic cell cloning technology, GTKO / β4GalNT2KO / CMAHKO / hCD39 / hCD46 / hCD55 / hCD59 / hTBM eight-gene-edited cloned pigs are constructed. Further, through genotype, mRNA, protein expression identification and functional analysis, eight-gene-edited xenogeneic organ transplantation donor pigs are obtained. The present invention solves the technical problems of high production difficulty, low efficiency, and low survival rate of donor pigs for multi-gene-edited xenogeneic organ transplantation, and maximally solves the common problems of immune rejection reaction and complement dysregulation faced during xenogeneic organ transplantation, laying a foundation for more targeted development of donor pigs suitable for different tissue and organ xenotransplantation, and having important value for promoting the clinical transformation of xenogeneic organ transplantation.

The invention discloses a micromolecule NSD3 degradation agent as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The invention provides a micromolecular NSD3 degradation agent, which has a structure as shown in a formula I or a formula II, has a remarkable inhibition effect on NSD3 protein expression, and has remarkable killing and growth inhibition effects on lung squamous carcinoma cells. The small molecule NSD3 degradation agent provides a new scheme for treatment of lung squamous cell carcinoma, and has a good clinical application prospect. # imgabs0 # imgabs1 #

The invention belongs to the technical field of living cell imaging, and discloses a living cell nucleic acid imaging system and method based on CRISPR (clustered regularly interspaced short palindromic repeats) and a nano antibody and application, the system comprises: a modified dCas protein expression vector containing n antigen polypeptide tags; the modified sgRNA and / or crRNA expression vector comprises a specific guide sequence; the fluorescent nano-antibody expression vector contains m fluorescent nano-antibody coding sequences which are connected in an operable manner, and fluorescent protein serving as an internal tag is inserted between nano-antibody splitting sites, can be combined with a polypeptide tag carried by dCas protein and can stably emit light; and the fluorescent nano antibody in a free state can be self-degraded, so that the fluorescent nano antibody does not emit fluorescence. According to the invention, the single-copy nucleic acid sequence, especially the imaging marker of the single-copy nucleic acid sequence in a living cell, can be visualized with single-molecule resolution and high signal-to-noise ratio.

The invention provides a breast cancer HER2 immunohistochemical digital slice automatic interpretation method, device and equipment and a storage medium. Relates to the technical field of digital pathological image analysis. Acquiring a full-slice image, automatically identifying an external contrast area and a tissue area through a multi-task head real-time instance segmentation model, and segmenting an ROI (Region of Interest) area of tumor cell aggregation; extracting image blocks by adopting a fixed sliding window based on the ROI region, and constructing a double-branch multi-task feature extraction model containing a backbone network: realizing HER2 expression pattern classification by an image block classification head, and outputting a continuous protein expression intensity quantized value by a dyeing intensity numerical head through regression analysis; and establishing a slice-level interpretation model, integrating the classification results and the intensity quantized values of all the image blocks, and outputting an automatic HER2 interpretation result. According to the invention, end-to-end analysis from cell-level characteristic quantification to slice-level diagnosis is realized, and an objective and quantifiable intelligent interpretation scheme is provided for HER2 immunohistochemical evaluation.

The invention provides a respiratory syncytial virus antigenic polypeptide, nucleic acid and a vaccine. Compared with the respiratory syncytial virus wild type Pre-F protein, the respiratory syncytial virus antigenic polypeptide or the immunogenic fragment of the respiratory syncytial virus antigenic polypeptide has the following changes: the 104th to 144th regions of the Pre-F protein are replaced with GS linker; two groups of disulfide bonds are introduced into S155C, S290C, A149C and Y458C; the S190F and the V207L are used for realizing cavity filling; and P102A, L373R, I379V, M447V, P102A, L373R, the C terminal of the respiratory syncytial virus antigenic polypeptide or the immunogenic fragment thereof comprises a cytoplasm tail region fragment, and the cytoplasm tail region fragment is a fragment derived from 3-10 continuous amino acid residues of a PreF protein cytoplasm tail region. According to the present invention, the pre-F immunogenicity and / or stability can be improved, and the protein expression level can be easily improved.

The present invention provides a yeast recombinant type XVII humanized collagen and its preparation method, in particular, a recombinant humanized collagen expressing the 15th helical region of type XVII collagen, a recombinant engineering bacterium and its preparation method, belonging to the technical fields of genetic engineering and synthetic biology. The present invention provides the full-length sequence of the 15th helical region of type XVII collagen, and mutants are obtained by mutating amino acid residues based on this sequence; for the first time, the present invention expresses the full-length sequence of the 15th helical region of type XVII collagen in Pichia pastoris, establishes a complete set of fermentation processes and purification processes, improves the protein expression level and reduces degradation, and through experiments, it is verified that the obtained protein has biological activities such as cell adhesion activity and cell migration promotion activity that reach or exceed those of natural human collagen, can achieve the purpose of real product application, and can be widely applied in the fields of medicine, medical devices, biological materials, tissue engineering, cosmetics, etc.

Provided are a respiratory syncytial virus antigenic polypeptide, a vaccine, and a use. First, a respiratory syncytial virus antigenic polypeptide or an immunogenic fragment thereof is provided. Compared with the amino acid sequence of a respiratory syncytial virus wild-type pre-F protein, the amino acid sequence of the respiratory syncytial virus antigenic polypeptide or the immunogenic fragment thereof has one or more mutation sites of amino acid residues at the following positions: positions 174-176, positions 181-184, positions 210-213, and positions 160-161. The improvement of pre-F immunogenicity can be achieved and / or the protein expression level can be improved.

The invention provides a medicine composition for treating cancer. The medicine composition comprises an inhibitor for inhibiting FOXRED2 protein expression and application of Paraprevir in preparation of a medicine for treating cancer. According to the application disclosed by the invention, the new indication of the hepatitis C medicine paritaprevir for inhibiting the malignant process of the liver cancer HCC is found for the first time, the treatment field of the medicine in the liver cancer is expanded, and a theoretical basis is provided for developing a new target spot for treating the liver cancer.

The invention belongs to the field of bioengineering, and particularly relates to phosphorylcholine cytidyltransferase and application thereof in preparation of citicoline, and the specific application is that the phosphorylcholine cytidyltransferase can efficiently catalyze phosphorylcholine and CTP to react to generate citicoline. The phosphorylcholine cytidyltransferase is prepared by the following steps: synthesizing a coding gene of the phosphorylcholine cytidyltransferase, selecting pHT01 as an expression vector, co-expressing with choline kinase, transforming into host bacteria, and inducing protein expression. According to the choline phosphate cytidyltransferase disclosed by the invention, under the same reaction condition and within the same reaction time, the yield of a citicoline product obtained by conversion is higher than that of choline phosphate cytidyltransferase (ScCCT) which is most commonly used in the market at present and is derived from saccharomyces cerevisiae S288c. Specifically, the yield of the MsCCT001 is 10 times higher than that of the ScCCT, the yield of the MbCCT006 is 6 times higher than that of the ScCCT, and the yield of the MeCCT008 is 8 times higher than that of the ScCCT.

Provided is a siRNA for inhibiting the gene expression of lfTSLP, said siRNA comprising a sense strand and an antisense strand. The provided siRNA can specifically target the long isoform of TSLP having pro-inflammatory activity, and inhibit the mRNA and protein expression of the long isoform of TSLP, while not affecting the mRNA and protein expression of the short isoform of TSLP having anti-inflammatory activity. Therefore, the siRNA has important clinical value for the treatment of lfTSLP related diseases.

The present disclosure relates to the seminal discovery of a generation and use of genetically engineered Vibrio sp. Provided is the use of the genetically engineered bacteria for the construction, maintenance, manipulation, and / or propagation of DNA constructs; protein expression; protein secretion; vectors and other metabolic tools; metabolic engineering; expression of cellular extracts for cell-free biology; shuttle vectors; cloning vectors; and for synthetic biology applications. The disclosure also relates to the use of the replication machinery of Vibrio sp. as a cloning or expression vector for replication of recombinant DNA constructs. The disclosure also relates to methods of use of the above.

The invention relates to application of 12-ketolithocholic acid in preparation of a medicine for preventing or treating acute ischemic intestinal injury. According to the application, 12-ketolithocholic acid (12-KLCA) is used for preventing or treating acute ischemic intestinal injury and is verified on a constructed classical intestinal ischemia reperfusion model, and a verification result shows that the 12-KLCA obviously improves intestinal histopathologic injury induced by intestinal ischemia reperfusion of a mouse, improves the survival rate of the mouse, and has a good application prospect in prevention or treatment of acute ischemic intestinal injury. The protein expression level of the apoptosis factor activated caspase-3 is reduced, the effect is obvious, safety and non-toxicity are achieved, and the side effect is small.

Provided are formulations, compositions and methods for delivering biological moieties such as modified nucleic acids into cells to modulate protein expression. Such compositions and methods include the delivery of biological moieties, and are useful for production of proteins.

The invention discloses a site-specific integration site screening method, a carrier particle, a site-specific integration method and a particle. The invention provides a site-specific integration site screening method, a carrier particle, a site-specific integration method and a particle, and aims to integrate upstream and downstream marker genes with recognition sequences on the carrier particle, perform protein expression quantity screening while performing monoclonal screening, obtain a monoclonal carrier with good performance through one-step screening, and perform site-specific integration on the carrier particle. The marker gene is replaced when the exogenous target gene is expressed, so that the overall stability of the particle is greatly ensured, the overall expression performance and stability of the expression particle are ensured while the development of the expression particle is shortened, and the technical problem that an expression vector in the prior art is relatively long in development time or poor in performance is solved. The site-specific integration site screening method provided by the invention can effectively shorten the development period of the expression vector with high expression quantity and overall stability. The carrier particle provided by the invention is good in adaptability and good in expression quantity and passage stability.