580 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 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.

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 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 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 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 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 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 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 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.

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 invention provides a streptococcus suis vaccine recombinant fusion protein and a preparation method and application thereof, the recombinant fusion protein is a recombinant protein formed by series fusion of B cell epitopes of Sao, SLY and PepO proteins of streptococcus suis, and the amino acid sequence of the fusion protein is as shown in SEQ ID NO.1. After four-week-old weaned piglets are immunized by the fusion expression protein for 28 days, 80% and 60% of challenge protection rates can be provided for challenge of streptococcus suis type 2 and type 9 virulent strains respectively. The invention has great potential for clinical prevention and control of streptococcus suis infection.

This invention relates to the field of genetic engineering technology, and provides a ZmPHYLL gene, the full-length nucleotide sequence of which is shown in SEQ ID NO: 1, and the coding region nucleotide sequence of which is shown in SEQ ID NO: 2. This invention also provides the application of the above-mentioned ZmPHYLL gene in inhibiting the germination of *Russula multifiliis* spores and improving the resistance of maize plants to southern maize rust. Simultaneously, this invention provides a method for improving the resistance of maize plants to southern maize rust. In this study, a ZmPHYLL gene was screened, and the expressed protein of this gene can effectively inhibit the germination of *Russula multifiliis* spores. Therefore, by regulating the overexpression of this gene, the resistance of maize to southern maize rust can be improved. This invention provides new gene resources for the creation of breeding materials and provides important scientific basis for the breeding of maize varieties highly resistant to southern maize rust.

The invention belongs to the field of biotechnology and biomedicine, and discloses an AAV system suitable for PFIC3 gene therapy and a preparation method. The system is composed of two adeno-associated virus vectors. The packaging plasmid of the adeno-associated virus vector comprises an auxiliary packaging plasmid pDP8.ape containing an AAV2Rep gene and an AAV8Cap gene, and a vector plasmid pAB288-Alb-hABCB4 for expressing an ABCB4 gene of human MDR3 protein, a self-cleavage protein P2A, a mouse albumin gene homologous arm sequence and an ITR sequence of AAV2, the packaging plasmid of the other adeno-associated virus vector comprises an auxiliary packaging plasmid pDP8.ape containing an AAV2Rep gene and an AAV8Cap gene, and a vector plasmid pX602-saCas9-gRNA for expressing a saCas9 protein and a sgRNA of a targeted mouse albumin genome.

The present application discloses an antibody drug conjugate, which comprises the structure of the following formula 1 wherein, Ab is an antibody or antibody fragment which targets ITGA2, and j is 1 to 8, preferably 4 to 8. The antibody-drug conjugate of the application has a high specificity binding ability to tumor cells expressing ITGA2 protein, thereby achieving an excellent killing effect on tumor cells.

The invention discloses an NDRV [sigma] C circular RNA vaccine and application thereof, and belongs to the technical field of vaccines.Duck hepatitis A virus type 1 (DHAV-1) UTR and T4 bacteriophage I type intron self-splicing cyclization systems are adopted to construct circular RNA (CirRNA-[sigma] C) for expressing NDRV [sigma] C protein, and a vaccine preparation is prepared through a chitosan-polyethyleneimine (CS-PEI) nano delivery system. The feasibility of a circular RNA vaccine system based on DHAV-1UTR in poultry vaccines is confirmed for the first time, the established nano delivery technology and circular RNA vaccine platform have the characteristics of high stability, good safety, strong immunogenicity and the like, and an important technical path is provided for development of novel vaccines of NDRV and other viral epidemic diseases.

The invention relates to a PagNRT3.1 gene for regulating xylem development of poplar and application of the PagNRT3.1 gene, and belongs to the technical field of plant genetic engineering and biology. The nucleotide sequence of the PagNRT3.1 gene for regulating and controlling the xylem development of the poplar is shown as a sequence 1 in a sequence table; the amino acid sequence of the expression protein of the PagNRT3.1 gene is shown as the sequence 2 in the sequence table. The PagNRT3.1 gene is transferred into poplar 84K, and compared with a wild type, the transgenic poplar over-expressing the PagNRT3.1 has a phenotype with increased xylem width, which indicates that the PagNRT3.1 gene is the key for regulating and controlling the development of the xylem of the poplar and has important application value in the field of high-yield genetic engineering of forest trees.

The invention belongs to the field of biological medicine, and provides circular RNA for treating multiple endocrine tumor type I and application of the circular RNA. According to the invention, after different optimization and design are carried out on the circular RNA for coding Menin, the circular RNA with relatively good expression level and long half-life period is finally obtained, and the anti-tumor effect is mainly achieved by supplementing cell-deficient Menin. Compared with the prior art, the circular RNA for expressing the Menin protein provided by the invention is more stable in intracellular expression and longer in half-life period. In an immortalized parathyroid gland cell model, the Menin protein can be expressed by transfecting the circular RNA for 6 hours, and the circular RNA has good treatment potential for multiple endocrine tumor type I.

The invention discloses a liriodendron hybrids LhCOLD1 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 LhCOLD1 gene disclosed by the invention is shown as SEQ ID NO. 1, and the amino acid sequence of the expression protein of the LhCOLD1 gene is shown as SEQ ID NO. 2. The method comprises the following steps: constructing an overexpression vector of a hybrid liriodendron LhCOLD1 gene, and transforming the overexpression vector into arabidopsis thaliana and hybrid liriodendron calluses; and cultivating and screening to obtain a transgenic line with improved low-temperature stress resistance. After the LhCOLD1 transgenic arabidopsis thaliana plant is subjected to low-temperature treatment at-10 DEG C for 3 hours and the LhCOLD1 transgenic arabidopsis thaliana plant recovers to grow at room temperature for two weeks, the growth of the LhCOLD1 transgenic After the LhCOLD1 transgenic hybrid liriodendron tulipifera strain is treated at the low temperature of-10 DEG C for 1.5 hours, the influence of low-temperature stress on the growth is small. The result shows that the LhCOLD1 gene enhances the tolerance of the plant to low-temperature stress.

The invention belongs to the technical field of genes, and discloses a method for efficiently expressing foreign protein based on an NC048599.1 site in a CHO cell genome. A site for stably expressing a protein in a CHO cell genome is located in a range of 69195000 to 69199000 of a CHO cell gene NC048599.1 with a sequence as shown in SEQ ID NO: 1 (Sequence Identifier Number 1), and the sequence of the CHO cell gene NC048599.1 is shown in SEQ ID NO: 1. Different protein genes are introduced at fixed positions in a CHO cell genome, and stable expression is carried out.

The invention belongs to the field of biological detection, and particularly relates to a kit for detecting the content of exosome CD82 protein. The kit comprises a standard substance, an elisa plate coated with a CA1 antibody, a CA2 antibody working solution marked by HRP, a buffer solution, a developing solution and a reaction stop solution. The kit provided by the invention can specifically detect the content of the CD82 protein in the blood exosome, has the advantages of simple and convenient detection method, high accuracy, good stability and the like as an auxiliary diagnosis means for gastric cancer with high content of the CD82 protein, and has a good application prospect.

The invention provides a subunit fusion protein tS on the surface of porcine D-type coronavirus as well as a preparation method and application of the subunit fusion protein tS, the preparation method comprises the following steps: 1) cloning a gene sequence as shown in SEQ ID NO.5 into an eukaryotic expression vector to obtain a recombinant plasmid containing a fusion protein tS coding gene; 2) transfecting a recombinant plasmid containing the fusion protein tS 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 S. The S can be expressed in a large amount in a soluble manner, the protein is stable, the problems that the S protein on the surface of the porcine D-type coronavirus cannot be expressed in a large scale and the like in the prior art are solved, and the preparation method is simple and low in cost.

The invention provides a novel therapeutic drug for treating Prom1 related retinal diseases. Specifically, the invention provides an optimized Prom1 gene expression cassette, an rAAV virus vector and a gene therapy drug. The medicine provided by the invention can specifically express the PROM1 protein in a retina photoreceptor layer, and is suitable for clinical application to treatment of Prom1 gene mutation related retinal diseases.

The invention belongs to the technical field of biology, and discloses a method for screening plant interaction protein by combining big data with artificial intelligence and verifying protein interaction by using yeast two-hybrid. According to the method, AI is combined with big data, and co-expression protein of family protein is obtained through big data screening and analysis; aI interaction prediction and molecular docking are carried out on the proteins, and interaction information is further analyzed and extracted. And further analyzing the condition of the screened interaction protein by using a yeast two-hybridization or fluorescence co-localization experiment. According to the method, aiming at the characteristics of complex genomes and abundant sequencing data of species such as wheat, public sequencing data is creatively utilized to carry out an interaction protein screening process of subgenomes. The problems of long protein screening period, high cost, large workload, false positive protein screening, no co-expression and the like are solved. The interaction relationship between important gene families can be quickly and conveniently obtained, the interaction structural domain can be accurately and clearly found, and technical support is provided for research on analysis of a wheat key gene functional mechanism.