773 results about "Expression protein" patented technology

The invention discloses a site for stably expressing protein in a CHO cell gene NW023276806.1 and application of the site, and belongs to the technical field of biological genes. The site belongs to a fixed position in a CHO cell genome, different protein genes are introduced based on a micro-homologous end connection mechanism through a CRISPR / Cas9 tool, and stable expression is carried out. By adopting a site-specific integration method, a target gene is integrated to a stable expression area in a site-specific manner, repeated high-expression monoclonal screening is effectively avoided, and an MMEJ mechanism is introduced to integrate a donor fragment, so that the research and development time for constructing a stable expression cell strain in biological pharmacy can be effectively shortened, and the cost is reduced.

The invention discloses a glycosyl transferase mutant and an application of the glycosyl transferase mutant in synthesis of rebaudioside. According to the invention, single-point mutation and multi-point mutation are carried out on the basis of a glycosyl transferase amino acid sequence as shown in SEQ ID NO: 1, and a mutant with improved catalytic activity is obtained. The catalytic activity, the substrate specificity and / or the substrate specificity of the glycosyl transferase mutant are / is changed, and the catalytic activity of enzyme to a specific substrate can be remarkably improved by mutation at a specific site. And carrying out induced expression and protein purification on the obtained mutation sequence to obtain the mutant enzyme. The mutant enzyme is used as a catalyst, and UDPG is used as a glycosyl donor, so that the catalytic reaction efficiency of substrates such as stevioside ST, rebaudioside A (RebA) and rebaudioside D (RebD) can be obviously improved. According to the glycosyl transferase UGT76G1 mutant constructed by the research, the catalytic activity of the glycosyl transferase UGT76G1 mutant is improved, and efficient production of rebaudioside M is realized by optimizing a reaction system.

The invention discloses a Candida antarctica lipase B mutant with high transesterification activity and application thereof. The nucleotide sequence of the Candida antarctica lipase B mutant is shown as SEQ ID NO.1, and the amino acid sequence of the expression protein of the Candida antarctica lipase B mutant is shown as SEQ ID NO.2. The invention further discloses a preparation method of the Candida antarctica lipase B mutant. Compared with an original enzyme, the mutant has higher fat hydrolysis activity and transesterification activity, the specific enzyme activity of the fat hydrolysis activity reaches 480.39 U / mg, the specific enzyme activity of the transesterification activity reaches 40.67 U / mg, the biodiesel conversion rate at the temperature of 30 DEG C can reach 92%, low-temperature high catalytic activity is shown, and the mutant can be used for enzymatic preparation of biodiesel and has potential industrial application value.

The invention discloses a modified bordetella pertussis strain and an application thereof. The modified Bordetella pertussis strain is characterized in that in the Bordetella pertussis strain BAA-589, the BP2992 protein is not expressed, or the inactive BP2992 protein is expressed, the pldA protein is not expressed, or the inactive pldA protein is expressed, and the mla F protein is not expressed, or the inactive mla F protein is expressed. When the Bordetella pertussis strain is treated by combining an ultrasonic method, compared with wild Bordetella pertussis BAA-589, the Bordetella pertussis strain has a remarkable high-foaming phenotype, the OMV secretion amount is remarkably increased, and a good technical foundation is laid for developing a Bordetella pertussis OMV vaccine.

The invention belongs to the technical field of bioengineering, and particularly relates to recombinant III-type humanized collagen and application thereof. Area fragments with good hydrophilicity in a human III-type collagen alpha chain are selected and sequentially connected to serve as expression protein, the amino acid sequence is 100% identical to that of a corresponding part of natural human III-type collagen, and immunological rejection cannot be generated when the human III-type collagen is applied to a human body. According to the invention, the screened new III-type humanized collagen sequence is transferred into pichia pastoris for recombinant expression, the prepared recombinant III-type humanized collagen has the advantages of strong stability, high expression quantity and good hydrophilicity, and meanwhile, the recombinant III-type humanized collagen also has obvious effects of promoting cell proliferation and cell adhesion.

A technique related to codon optimization is provided. In a method for optimizing nucleic acid sequences expressing proteins in a host, protein subsequence-nucleic acid subsequence pairs are obtained from collected highly expressed protein sequences and their encoding nucleic acid sequences to form a training set. And training a neural machine translation model by using the training set. The neural machine translation model realizes translation from an amino acid sequence to a codon sequence. A protein sequence needing codon optimization is cut into protein subsequences. And translating a protein subsequence from an amino acid sequence to a codon sequence by using the trained neural machine translation model. The translated subsequences are superimposed to merge into a full-length codon sequence. In the process of superimposing to synthesize the codon sequence, according to the frequency or number of synonymous codons corresponding to each amino acid position of the protein sequence, the synonymous codon with the highest occurrence frequency or number is selected as the optimal codon at the position.

The invention relates to a mulberry MaBEH1 / 2 gene for improving plant salt tolerance and application thereof, and belongs to the technical field of plant genetic engineering and biology. Wherein the coding nucleotide sequence of the mulberry MaBEH1 / 2 gene for regulating and controlling the salt tolerance of the woody plant is shown as a sequence 3; and the amino acid sequence of the MaBEH1 / 2 expression protein is shown as a sequence 4. According to the invention, MaBEH1 / 2 is transferred into 84K poplars to obtain MaBEH1 / 2-OE transgenic poplars; compared with a non-transgenic poplar, the salt stress tolerance of the MaBEH1 / 2-OE transgenic poplar can be remarkably enhanced. The invention provides key gene resources and technical support for genetic improvement of salt-tolerant forest varieties.

The invention discloses a chicken infectious anemia virus subunit vaccine, and belongs to the technical field of animal disease vaccines. The invention provides a chicken infectious anemia virus subunit vaccine. Active ingredients of the chicken infectious anemia virus subunit vaccine are recombinant fusion protein of chicken infectious anemia virus VP1 protein B cell epitope and salmonella typhimurium flagellin. The B cell epitope of the chicken infectious anemia virus VP1 protein is obtained by connecting B cell antigen epitopes contained in the chicken infectious anemia virus VP1 protein through a connecting peptide. The salmonella typhimurium flagellin is obtained by deleting a hypervariable region of the salmonella typhimurium flagellin FliC. The fusion expression protein provided by the invention can induce generation of a specific IgG antibody after one-day-old SPF chicken, and after immunization for 28 days, immunized chicken flocks can resist challenge of chicken infectious anemia virus virulent strains, and 70% of challenge chicken flocks are protected without disease attack. The invention has great potential for clinical prevention and control of chicken infectious anemia virus infection.

The invention belongs to the technical field of biological medicine, and particularly relates to a kit for breast cancer diagnosis and application thereof. The kit comprises an elisa plate coated with a captured antibody and an HRP labeled antibody working solution, the capture antibody is a monoclonal antibody T2X-1 of an anti-PSTPIP1 protein; the HRP labeled antibody is a monoclonal antibody Z4Y-2 of an anti-PSTPIP1 protein, and the HRP labeled antibody is a monoclonal antibody Z4Y-2 of an The heavy chain amino acid sequence of the monoclonal antibody T2X-1 is as shown in SEQ ID NO.1, and the light chain amino acid sequence of the monoclonal antibody T2X-1 is as shown in SEQ ID NO.2; the heavy chain amino acid sequence of the monoclonal antibody Z4Y-2 is as shown in SEQ ID NO.3, and the light chain amino acid sequence of the monoclonal antibody Z4Y-2 is as shown in SEQ ID NO.4. The kit for breast cancer diagnosis provided by the invention can be used as an auxiliary diagnosis means for breast cancer with high expression of PSTPIP1 protein, and has high diagnostic value.

The invention discloses a poplar PdbHLH57 gene as well as an expression protein and application thereof, and relates to the technical field of plant genetic engineering. The nucleotide sequence of the poplar PdbHLH57 gene disclosed by the invention is as shown in SEQ ID NO.1, and the amino acid sequence of the expression protein of the poplar PdbHLH57 gene is as shown in SEQ ID NO.2. The method comprises the following steps: constructing an expression vector of a poplar PdbHLH57 gene, and transforming the expression vector into a poplar leaf; and cultivating and screening to obtain the transgenic poplar with red leaves. Results show that compared with wild poplar, the PdbHLH57 gene for promoting poplar anthocyanin synthesis has the advantages that transgenic poplar leaves become red, and the anthocyanin content in transgenic plant leaves is remarkably increased; the expression level of anthocyanin biosynthesis related genes is remarkably increased, and further, it is shown that the PdbHLH57 gene plays an important role in poplar anthocyanin biosynthesis.

The invention belongs to the field of enzyme engineering, and relates to a glycosyl transferase mutant and application thereof in synthesis of salidroside. On the basis of a glycosyl transferase amino acid sequence as shown in SEQ ID NO: 1, iterative saturated mutation is carried out through a molecular modification technology, and the enzymatic activity and catalytic efficiency of the modified glycosyl transferase mutant are greatly improved. The mutation at a specific site can significantly promote the glycosylation activity of a tyrosol substrate. The mutant enzyme is obtained by carrying out induced expression and protein purification on the obtained mutant strain. The substrate conversion rate of the transformed glycosyl transferase mutant for catalyzing the tyrosol glycosylation reaction can reach 95% or above, the yield reaches 55.0 g / L, and an efficient and green new way is provided for salidroside production.

The invention provides a subunit fusion protein RBD-Fc on the surface of a porcine D-type coronavirus and a preparation method and application thereof, the preparation method comprises the following steps: 1) cloning a gene sequence as shown in SEQ ID NO.5 into a eukaryotic expression vector to obtain a recombinant plasmid containing a fusion protein RBD-Fc coding gene; (2) transfecting a recombinant plasmid containing the fusion protein RBD-Fc coding gene into an expression cell; 3) culturing, screening and domesticating the expression cells in the step 2) to obtain a highly expressed cell strain; and 4) fermenting and culturing the cell strain in the step 3), and purifying to obtain the subunit fusion protein RBD-Fc. According to the present invention, the RBD-Fc can be subjected to mass soluble expression, the protein is stable, the problems that the porcine D-coronavirus surface S protein cannot be subjected to large-scale expression and the like in the prior art are solved, the preparation method is simple, and the cost is low.

The present invention relates to pluripotent stem cell comprising an expression construct for expression of a CEBPα protein and an expression construct for expression of an ADIG protein. The invention further provides for methods of producing adipocytes comprising the pluripotent stem cells and for foodstuff comprising the adipocytes or pluripotent stem cells.

The invention belongs to the technical field of genes, and discloses a method for efficiently expressing a foreign protein based on an NC048604-1 site in a CHO cell genome. A site for stably expressing protein in the CHO cell genome is located in the 94142000 to 94148000 basic group range of the CHO cell genome NC048604-1, and the nucleotide sequence of the site is as shown in SEQ ID NO: 1. According to the invention, different protein genes are introduced at fixed positions in a CHO cell genome, and stable expression is carried out.

The invention discloses a chimeric PEDV (porcine epidemic diarrhea virus) infectious clone plasmid, a PEDV chimeric virus as well as a construction method and application thereof. According to the invention, a PEDV genome is used as a skeleton for the first time, the chimeric virus for expressing the extracellular domain of the TGEV S protein is successfully rescued, and it is found that the TCID50 of the chimeric virus for expressing the extracellular domain of the TGEV S protein can be increased by 6.2 times compared with that of the chimeric PEDV for expressing the TGEV S protein. The recombinant virus is used as an inactivated vaccine for immunizing mice, so that the immunogenicity of TGEV S protein can be greatly improved, a body is stimulated to generate a higher-level neutralizing antibody aiming at TGEV, and the titer of the neutralizing antibody aiming at TGEV is improved by 16.8 times. Therefore, more possibilities are provided for modification of viral genomes, a novel method for improving immunogenicity of the TGEV inactivated vaccine is invented, and a basis is provided for research and development of the TGEV vaccine.

The invention discloses a method for inducing plant disease resistance by using a hydrolase active mutant of xyloglucan specific glucanase XEG1. A pichia pastoris eukaryotic expression system is utilized to obtain the purified hydrolase activity mutant XEG1E136D protein of the phytophthora sojae core pathogenic factor xyloglucan specific glucanase XEG1, and the hydrolase activity mutant XEG1E136D protein does not have the hydrolase activity of plant cell walls and can activate the systemic resistance of plants. The XEG1E136D is overexpressed into a biocontrol bacterium trichoderma harzianum by utilizing an agrobacterium-mediated fungal transformation optimization system, and the XEG1E136D is proved to be capable of secreting XEG1E136D protein to the outside of cells and activating the broad-spectrum systemic resistance of plants. The XEG1E136D protein expressed by the trichoderma harzianum can be used as a plant vaccine to improve the prevention and treatment effect, and the trichoderma harzianum XEG1E136D protein has a wide application prospect in the field of plant disease prevention and treatment.

The present invention provides a novel therapeutic drug for treating a Prom1-associated retinal disease. Specifically, the present invention provides an optimized Prom1 gene expression cassette, an rAAV viral vector, and a gene therapy drug. The drug of the present invention can specifically express the PROM1 protein in the retinal photoreceptor layer, and is suitable for the clinical treatment of a retinal disease associated with Prom1 gene mutation.

The invention discloses a mint McTTG2 gene as well as an expression protein and application thereof, and relates to the technical field of plant genetic engineering. The nucleotide sequence of the mint McTTG2 gene disclosed by the invention is shown as SEQ ID NO. 1, and the amino acid sequence of the expression protein of the mint McTTG2 gene is shown as SEQ ID NO. 2. The method comprises the following steps: constructing an expression vector of a mint McTTG2 gene, and transforming the expression vector into an arabidopsis thaliana mutant; and cultivating and screening to obtain a transgenic arabidopsis thaliana plant of which the procyanidine content is obviously increased. A transgenic experiment shows that after the McTTG2 gene is over-expressed in the mutant ttg2-7, the content of procyanidine is remarkably increased; compared with the mutant ttg2-7, the number of epidermal hairs on leaves of ttg2-7 / 35S:: McTTG2-1 and-2 strains is obviously increased, which is close to that of wild strains. Gene resources are provided for plant variety improvement, and cultivation of new mint varieties is facilitated.

The invention provides a glomerular senescence evaluation method, system and device, and relates to the technical field of glomerular senescence evaluation.The evaluation method mainly comprises the steps that based on a low-abundance proteomics result of an aged mouse plasma sample, core expression protein related to glomerular senescence is screened out, a prediction model aiming at glomerular senescence is constructed, and the prediction model is used for predicting the glomerular senescence; the prediction model comprises a glomerular senescence index; detecting core expression protein data of a to-be-detected plasma sample, and calculating the glomerular senescence index of the to-be-detected plasma sample according to the prediction model. According to the scheme, aiming at the specificity of glomerular senescence, core expression protein can be screened out and a prediction model can be constructed through methods such as low-abundance proteomics results, so that the glomerular senescence can be evaluated simply, conveniently, reliably and quickly; the scheme is helpful for follow-up efficient research and evaluation of anti-glomerular senescence drugs, and is helpful for follow-up efficient research and explanation of the effect of glomerular senescence in CKD progress.

The invention relates to the technical field of molecular biology, in particular to a multichannel virus reporting system based on TAT trans-activation. The invention relates to a multichannel virus reporting system based on TAT trans-activation. The multichannel virus reporting system comprises an activated virus and a reporting cell, the activated virus is a human enterovirus A71 for expressing TAT protein; the reporter cells are cells transfected with reporter plasmids; the reporter plasmid comprises an LTR sequence and a plurality of reporter genes which are started to express by the LTR sequence. When the virus is activated to infect the reporter cell, the TAT protein carried by the virus enters the reporter cell to activate the expression of the downstream reporter gene, and the expression level of the reporter gene can reflect the virus replication condition, so that the screening or mechanism research of related antiviral drugs can be realized. According to the technical scheme, the technical problem that the flexibility, stability and high-throughput screening adaptability of a virus reporting system in the prior art are not ideal can be solved, and the virus reporting system has ideal popularization and application prospects.

The invention discloses a mint McTTG1 gene as well as an expression protein and application thereof, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the mint McTTG1 gene disclosed by the invention is shown as SEQ ID NO.1, the amino acid sequence of the expression protein of the mint McTTG1 gene is shown as SEQ ID NO.2, and the mint McTTG1 gene belongs to WD40 family genes participating in regulation and control of epidermal hair development and synthesis of anthocyanin and procyanidine. Transgenic experiments show that after the McTTG1 gene is over-expressed in the arabidopsis thaliana mutant ttg1-13 with epidermal hair loss and significantly reduced accumulation of anthocyanin and procyanidine, the formation of the epidermal hair on the surface of the mutant can be recovered, and the accumulation of anthocyanin and procyanidine can be promoted. Gene resources are provided for plant variety improvement, and cultivation of new mint varieties is facilitated.

The present invention relates in part to nucleic acids encoding proteins, nucleic acids containing non-canonical nucleotides, therapeutics comprising nucleic acids, methods, kits, and devices for inducing cells to express proteins, methods, kits, and devices for transfecting, gene editing, and reprogramming cells, and cells, organisms, and therapeutics produced using these methods, kits, and devices. Methods for inducing cells to express proteins and for reprogramming and gene-editing cells using RNA are disclosed. Methods for producing cells from patient samples, cells produced using these methods, and therapeutics comprising cells produced using these methods are also disclosed.

The invention belongs to the technical field of biology, and particularly relates to a kit for detecting breast cancer and a detection method. The kit comprises an elisa plate coated with a capture antibody L4-CAP, a biotin-labeled detection antibody L4-SIG, a protein standard substance, HRP, a sample diluent, a washing solution, a developing solution and a stop solution. The heavy chain amino acid sequence of the capture antibody L4-CAP is as shown in SEQ ID NO.1, and the light chain amino acid sequence of the capture antibody L4-CAP is as shown in SEQ ID NO.2; the heavy chain amino acid sequence of the detection antibody L4-SIG is shown as SEQ ID NO.3, and the light chain amino acid sequence of the detection antibody L4-SIG is shown as SEQ ID NO.4. The kit for detecting the breast cancer is high in specificity, can specifically detect the content of the URCL4 protein in serum, serves as an auxiliary diagnosis means for breast cancer with high expression of URCL4 protein, and has a good application prospect.

The invention discloses a 5 '-UTR sequence derivative based on SARS-CoV-2 and application of the 5'-UTR sequence derivative in enhancement of plant gene translation expression, and relates to the technical field of enhancement of plant gene translation expression. The 5 '-UTR sequence derivative provided by the invention is used for enhancing translation expression of a target gene in a plant body; the nucleotide sequence of the 5 '-UTR sequence derivative is as shown in SEQ ID NO. 1 or SEQ ID NO. 2. The SARS-CoV-2 virus 5 '-UTR is applied to a plant system for the first time, through the optimized 5'-UTR sequence derivative mediated translation regulation effect, the level of target gene expression protein is remarkably improved on the translation level, and the SARS-CoV-2 virus 5 '-UTR is suitable for various plant varieties such as gramineous plants, cruciferous plants and solanaceae plants.

The present invention relates to pluripotent stem cell comprising an expression construct for expression of a Myo1B protein. The invention further provides for methods of producing adipocytes comprising the pluripotent stem cells and for foodstuff comprising the adipocytes or pluripotent stem cells.

The invention discloses a dissolution-promoting expression tag and application thereof, and belongs to the technical field of gene engineering. The solubilizing expression tag is thioredoxin Trx1 or Trx2 from yeast, and the amino acid sequence of the solubilizing expression tag is as shown in SEQ ID NO. 1 or SEQ ID NO. 2. Compared with other solubilizing tags from biological sources, the solubilizing expression tag provided by the invention can better promote correct folding of the exogenous fusion protein in the yeast, improve the soluble expression quantity and overcome the condition that the soluble expression of the heterologous protein in a yeast expression system is not ideal. The solubilizing expression tag is applied to heterologous expression protein of a yeast expression system, expression of heterologous protein is provided, the yield of target metabolites is further increased, and the solubilizing expression tag has a good industrial application prospect.

The invention discloses an influenza C virus HEF protein polyclonal antibody preparation method, which comprises: carrying out signal peptide prediction, antigen epitope screening and hydrophobicity analysis through bioinformatics, constructing a recombinant prokaryotic expression vector pET32a-HEF by using a seamless cloning technology, and efficiently expressing the HEF protein in Escherichia coli to obtain the influenza C virus HEF protein polyclonal antibody. The inclusion body protein yield is improved by optimizing conditions; and purifying by nickel column affinity chromatography to obtain high-purity recombinant protein, emulsifying the high-purity recombinant protein and a Freund's adjuvant, immunizing a Japanese white rabbit, immunizing for three times, and collecting high-titer serum. The method breaks through detection limitation caused by antigen tag concealment in a eukaryotic expression system, the obtained polyclonal antibody is high in specificity, the eukaryotic expression HEF protein can be accurately recognized, and a key tool is provided for ICV virus-like particle identification, vaccine research and development and diagnosis technologies. Compared with a traditional method, the scheme is simple and convenient to operate, low in cost and high in antibody titer, and has remarkable application value.

The invention discloses a porcine delta-coronavirus double-antibody sandwich ELISA (enzyme-linked immuno sorbent assay) detection kit based on an S protein monoclonal antibody as well as a use method and application of the porcine delta-coronavirus double-antibody sandwich ELISA detection kit. A PDCoV S protein is expressed by adopting an ExpiCHO expression system, then the purified S protein is used as an immunogen to immunize a mouse, and the monoclonal antibody is obtained. And then, respectively using the monoclonal antibody 3D7 and the MAb 9G4 labeled by the HRP as capture and detection antibodies, and establishing the double-antibody sandwich ELISA kit. The kit has good sensitivity, the detection limit of purified S protein is 0.12 ng / mL, and the detection limit of PDCoV is 1.96 * 10 < 3 > copies / [mu] L. Specificity analysis shows that the kit has no cross reactivity with other porcine intestinal coronaviruses PEDV, TGEV and PoRV with clinical symptoms similar to PDCoV, the consistency with an RT-PCR (reverse transcription-polymerase chain reaction) method reaches 91.03%, and the Kappa value is 0.814, so that the reliability of the kit for clinical sample detection is shown. In addition, the kit can also quantitatively evaluate the content of S protein in PDCoV inactivated vaccines and subunit vaccines, and provides a powerful tool for evaluating the vaccines.

A recombinant interleukin-27 (IL27) expressing virus is described. The recombinant IL27 expressing virus comprises an oncolytic virus comprising one or more exogenous nucleic acid sequences capable of expressing in IL27 protein or a biologically active portion thereof, the exogenous nucleic acid sequences being operably linked to an expression control sequence. Methods of treating cancer by in a subject by contacting a cancer cell of the subject with a recombinant IL27 expressing virus are also described.

The present invention belongs to the field of biomedicine, and specifically relates to a new use of Keratin 7 protein, and a recombinant plasmid expressing Keratin 7 protein, a recombinant probiotic expressing Keratin 7 protein, and a use for preparing a medicine for treating and / or preventing metabolic diseases. The present invention first inserts the Keratin 7 gene into the expression vector pET‑28a to obtain a recombinant plasmid expressing Keratin 7; secondly, the recombinant plasmid is introduced into Escherichia coli Nissle 1917 (EcN) tolerant bacteria to obtain a functional recombinant probiotic. The recombinant probiotics described in the present invention can effectively reduce blood sugar while efficiently expressing Keratin 7 protein, and has a significant therapeutic effect on diabetes and its complications, and the effect is higher than that of clinical drugs, and has good clinical application prospects.