103 results about "Expression cassette" patented technology

This invention relates to the fields of bioengineering and fermentation engineering, and discloses a genetically engineered *Streptomyces albopictus* strain that produces high levels of ε-polylysine and its fermentation production process. This genetically engineered strain uses *Streptomyces albopictus* strains acclimated to ε-polylysine tolerance as a chassis, and its genome integrates... ppc and dapf Through expression box, heterogeneous asd Expression cassettes, containing secretory signal peptides lysp Fusion expression cassettes and D404A / K499A point mutations pls Expression cassette. The fermentation process employs a two-stage pH control strategy, maintaining a neutral environment during the cell growth phase and adjusting to an acidic environment during product synthesis. This invention improves ε-polylysine yield through a synergistic strategy of enhancing precursor supply, constructing efflux detoxification channels, and increasing synthase activity; combined with an acidic fermentation process, it effectively inhibits product degradation, yielding a high-molecular-weight and highly uniform target product suitable for industrial production.

This invention provides a high-throughput method for constructing dual sgRNA libraries and its applications. The method utilizes high-throughput microarray synthesis technology to prepare a set of DNA fragments containing multiple dual sgRNA expression cassettes in a single step. After amplification, these fragments are assembled with a target vector containing a first promoter and a second gRNA backbone sequence in a first round of directed assembly to obtain a preliminary recombinant plasmid set. Then, linearized enzyme digestion and homologous recombination technology are used to insert a fragment containing a transcription termination sequence and a complete second promoter to complete the construction of the dual sgRNA expression unit. Finally, transformation and amplification yield the dual sgRNA plasmid library. This invention avoids the high error rate and high cost of long-chain oligonucleotide synthesis by utilizing microarray synthesis and simplifies the operation process through two rounds of directed assembly, significantly improving the throughput, fidelity, and efficiency of library construction. It is applicable to the construction of genome-wide dual sgRNA libraries, providing an efficient and reliable technical platform for high-throughput gene function screening, drug target discovery, and gene interaction research based on CRISPR.

The application discloses a sorghum double-grain trait related gene DG1 and application thereof. The application provides application of any one of the following A1) to A3) in regulation of plant ear grain number, regulation of plant yield and / or cultivation of multiple-grain plants: A1), a protein DG1; A2), a nucleic acid molecule encoding the protein DG1; and A3), a recombinant carrier, an expression cassette or a recombinant bacterium containing the nucleic acid molecule encoding the protein DG1. The application finds that the protein DG1 is a gene for regulating plant ear grain number, overexpression of the gene can increase the plant ear grain number, make the sorghum multiple-grain and realize high yield.

This invention provides a tandem gene, expression cassette, expression vector, and rice germplasm construction method for synthesizing ginsenoside Ro, belonging to the field of genetic engineering technology. This invention provides a tandem gene for synthesizing ginsenoside Ro, which involves tandemly connecting four key genes from *Saccharomyces cerevisiae*, one gene from *Licoricea javanica*, and five genes from ginseng, and constructing gene expression units for each, thereby forming a tandem expression cassette. This invention connects the tandem expression cassette with a plant expression vector to construct a recombinant expression vector, transforms the recombinant expression vector into *Agrobacterium*, and successfully transforms the recombinant expression vector into the rice genome using an *Agrobacterium*-mediated genetic transformation method. The rice can express the recombinant expression vector correctly and efficiently in the grains, laying a solid technical foundation for the large-scale synthesis of ginsenoside Ro.

Provided herein are multivalent proteins comprising a first region capable of binding to one or more effector proteins and a second region capable of binding to one or more target proteins. Also provided are expression cassettes and vectors comprising nucleic acids encoding the multivalent proteins. Also provided are recombinant cells comprising the expression cassettes and vectors. Also provided are uses of such multivalent proteins, nucleic acids and recombinant cells. Also provided are the multivalent proteins, nucleic acids and recombinant cells for use as medicaments.

The application discloses a biomaterial related to DCLK1 and application of DCLK1 in reversing an immunosuppressive microenvironment after EGFR-TKI drug resistance of lung adenocarcinoma, and belongs to the technical field of biotechnology.The biomaterial is an RNA molecule for inhibiting or reducing or down-regulating expression of a coding gene of a protein, or an RNA molecule for inhibiting or reducing or down-regulating activity or content of the protein, or an RNA molecule for knocking out a coding gene of the protein, or a coding gene, an expression cassette and the like related to the foregoing RNA molecule.The application shows through experiments that targeting DCLK1 can inhibit a GAS6 / AXL pathway, and down-regulate expression of PD-L1 and IDO1, thereby recovering T cell infiltration and proliferation capacity, reversing an immunosuppressive microenvironment after drug resistance, and indirectly suppressing progression of lung adenocarcinoma with three generations of EGFR-TKI drug resistance.

Recombinant DNA molecules and constructs are provided that are useful for modulating gene expression in plants. One or more expression cassette(s) of a recombinant DNA molecule or construct may be excised from transgenic plants following transformation by the presence of flanking site-specific recombination sites in the recombinant DNA molecule or construct by expression of a recombinase enzyme encoded by the recombinant DNA molecule or construct. Such a recombinase system may be used to remove such expression cassette(s) from plants transformed with the recombinant DNA construct or vector. The recombinase transgene may be operably linked to a tissue-preferred or tissue-specific promoter for autoexcision in transformed plants without crossing to a different transgenic line expressing the recombinase. Methods for causing autoexcision of one or more expression cassette(s) in a transgenic plant, and plants and cells containing or transformed with a recombinant DNA molecule or construct of the present disclosure, are also provided.

This invention discloses a plasmid for gene overexpression in non-pattern *Bacillus laterosporus*. Its leader region contains a methyltransferase expression cassette derived from the *Bacillus laterosporus* i83 genome, enabling stable existence in specific hosts containing the RM system. Based on this, a PidR2 promoter is added to obtain the pMCPidR2 shuttle plasmid. The PidR2 promoter effectively drives target gene expression, the pWV01 replicon ensures plasmid replication in non-pattern hosts, and the chloramphenicol resistance gene is used for positive colony selection. The plasmid contains a TraJ element, allowing it to be introduced into non-pattern microbial hosts via conjugation transfer. This invention also provides a target gene expression strategy in non-pattern microorganisms, achieving high transformation efficiency and high strain yield through conjugation transfer transformation of donor bacteria, conjugation transfer helper bacteria, and recipient bacteria, resulting in stable expression of the target protein. This invention provides an effective vector for efficient and stable gene overexpression in specific non-pattern microorganisms, helping to solve the problem of difficult genetic manipulation of industrial production strains.

A vector set is introduced into a host cell, and a cell that produces a multispecific antibody is selected from the host cell into which the vector set has been introduced. The vector set is a set of a first expression vector and a second expression vector, in which the first expression vector contains an expression cassette of a first H chain, an expression cassette of a second H chain, an expression cassette of a first L chain, and an expression cassette of a second L chain, and the second expression vector contains one type, two types, or three types of expression cassettes selected from the group consisting of the expression cassette of the first H chain, the expression cassette of the second H chain, the expression cassette of the first L chain, and the expression cassette of the second L chain.

Methods and compositions are provided for enhancing the accumulation and / or recovery of recombinant casein expressed in soy. Endogenous Casein-Processing Interfering Proteins (CPIPs), such as oxidases like lipoxygenases (LOX), are present in the recombinant protein fractions and interfere with yield and / or purity. RNA interference (RNAi) mechanisms are leveraged in plasmids that knockdown genes expressing CPIPs and / or genes in the Soybean Vesicular Trafficking Pathway (SVTP). Knocking down of two or more target genes is manifested using a dual gene targeting approach, often via a single expression cassette containing multiple distinct shRNA constructs for enhanced efficiency. The shRNA constructs invoke a RNAi mechanism that leads to increased casein accumulation either: (1) directly, by suppressing CPIPs (e.g., LOX) to enable more efficient purification; or (2) indirectly, by modifying SVTP genes to route casein away from lytic vacuoles, preventing degradation.

This invention relates to the field of biomedical technology, specifically disclosing a gene modification method for enhancing TCR-T cell activity and its application in tumor treatment. The method includes the following steps: T cells are activated in vitro; gene editing is performed to knock out endogenous immune checkpoint genes; a recombinant expression vector is constructed, the vector containing a polycistronic expression cassette driven by a single promoter, wherein the nucleic acid sequence encoding TCR and the nucleic acid sequence encoding an immunomodulatory factor are tandemly linked by a self-cleaving peptide coding sequence; the recombinant expression vector is introduced into the gene-knockout T cells; and the transduced T cells are cultured and expanded to obtain TCR-T cells with enhanced activity. This invention significantly enhances the anti-tumor activity, in vivo persistence, and tumor infiltration capacity of TCR-T cells through the synergistic effect of knocking out immune checkpoint genes and autocrine immunomodulatory factors; its modular design allows for flexible adaptation to different tumor antigen targets.

The application discloses a method for improving cellulase activity by using a combination of an endogenous promoter and an associated 5' UTR of Trichoderma reesei and an engineering strain constructed by the method. TrAE The 5' UTR upstream of the TrAE gene TrAE Overexpressing a transcriptional regulator xyr1 The expression cassette of the gene is transformed into Trichoderma reesei Δ ku70 Rut‑C30 The application also discloses a high-yield cellulase Trichoderma reesei strain QEaeuX, and a method for producing high-activity cellulase by using the strain. ku70 Rut‑C30 The cellulase activity produced by the strain in a 96 h shake flask culture is 2.6 IU / mL, which is 101% higher than that of the original strain Δ The application has a good industrial development and application prospect.

The present invention relates to a closed circular double-stranded DNA molecule comprising a poly(dA) sequence from 25 to 200 nucleotide residues in length and an expression cassette, wherein the poly(dA) sequence is located at a position within the closed circular double-stranded DNA molecule such that, when the closed circular double-stranded DNA molecule is incubated under conditions suitable for transcription, the poly(dA) sequence is not transcribed; and when the closed circular double-stranded DNA molecule is incubated under conditions suitable for replication, the poly(dA) sequence does not interfere with the replication of the closed circular double-stranded DNA molecule. The invention also relates to a method for the isolation or purification of DNA molecules, to uses of poly(dA) and oligo(dT) in the isolation or purification of DNA molecules, and to isolation or purification kits.

The application discloses a kind of recombination of Pseudomonas putida for producing L-lactic acid using whole-component lignocellulose hydrolysate and its construction method and application, belong to the field of metabolic engineering.The application uses Pseudomonas putida KT2440 as chassis, realizes carbon flow to pyruvic acid convergence and prevents product degradation by knocking out aceEF 、 ppsA And lldD Gene;By in-situ replacement, introduce optimized Lactobacillus plantarum ldhL1 Reconfiguration L-lactic acid synthesis pathway;By integration xylD-yjhH Tandem expression cassette enables xylose utilization;And by quorum sensing promoter P Rox306 Overexpression maeB Realize funnel conversion of lignin monomer.The application innovatively uses lignin derivatives in hydrolysate as natural donor of endogenous acetyl-coenzyme A, makes up lethal growth defect caused by knocking out aceEF The engineering bacteria of the application realize the synchronous high-efficiency conversion of glucose, xylose and aromatic monomer under the condition of not adding exogenous auxiliary carbon source and chemical inducer, which greatly reduces the industrial fermentation cost.

This invention provides an expression cassette, an expression vector, a genetically engineered strain, and their application in the preparation of malonic acid. The expression cassette includes expression cassette 1, expression cassette 2, and expression cassette 3. Expression cassette 1 includes the gene for the mitochondrial 37S ribosomal protein mS47; expression cassette 2 includes the ACC1 gene, which incorporates a mitochondrial localization signal encoding gene at its 5' end; and expression cassette 3 includes the upstream homologous arm of an endogenous ACC1 gene, which incorporates a weak promoter at its 5' end. This invention, through the expression cassette, can promote an increase in the levels of mS47 and ACC1 in mitochondria, thereby promoting the synthesis pathway of acetyl-CoA-malonyl-CoA-malonic acid (ACEA), while simultaneously reducing the level of endogenous ACC1 in the cytoplasm, avoiding competitive effects on the mitochondrial ACEA synthesis pathway, and ultimately effectively increasing malonic acid production.

This invention relates to a method for creating late-flowering, high-yielding alfalfa, characterized by the following steps: S1: designing and synthesizing targeted alfalfa MvFT S1: Construct the gRNA of the gene; S2: Construct a CRISPR / Cas9 gene knockout vector containing the gRNA expression cassette described in S1; S3: Transform the knockout vector described in S2 into Agrobacterium tumefaciens; S4: Transform explants of alfalfa cultivars into Agrobacterium tumefaciens as described in S3; S5: Screen, differentiate, and regenerate the explants transformed in S4 to obtain transgenic plants; S6: Perform molecular and phenotypic identification on the transgenic plants to screen for... MvFT A late-flowering, high-yielding germplasm with homozygous gene knockout and significantly delayed flowering time. This invention reveals for the first time... MvFT The key functions of genes in the regulation of flowering in alfalfa were investigated, and new late-flowering, high-yielding alfalfa germplasm was successfully created, providing valuable germplasm resources for breeding new high-yielding and high-quality alfalfa varieties.

This invention belongs to the field of plant gene editing technology, specifically relating to a bifunctional gene editing system and its application in single-parent maize. The invention provides a bifunctional gene editing system comprising: a fusion protein expression cassette and a guide RNA combined expression cassette; wherein the guide RNA combined expression cassette includes: pegRNA associated with the maize D9 gene; and sgRNA associated with the maize YUC4 and YUC2 genes. Transient expression validation using the bifunctional gene editing system in maize protoplasts revealed that the system achieves a precise editing efficiency of 3.5% at the G571V site of the D9 gene, a knockout efficiency of 40%-65% for the YUC4 gene, and a knockout efficiency of 37%-54% for the YUC2 gene. This demonstrates that the technical solution of this application can simultaneously and efficiently achieve precise point mutation and gene knockout within the same cell, thus enabling its application in the synergistic improvement of lodging resistance in maize.

The present invention relates to compositions and methods for treating complement-mediated diseases. Provided are recombinant vectors having an expression cassette comprising a modified human factor H (hfH) gene, wherein the hfH gene encodes a hfH protein variant comprising SCR1-4, 19-20, and one or more of: SCR7, SCR17, and / or SCR18. Also provided are pharmaceutical compositions comprising the vectors and their use in treating AMD and / or other complement-related diseases.

This invention relates to gene constructs, expression cassettes comprising said gene constructs, and oncolytic viruses, and their use in methods for treating, preventing, or alleviating cancer. The gene constructs, expression cassettes, and oncolytic viruses are particularly useful (but not limited to) for treating bladder cancer, such as non-muscle-invasive bladder cancer.

Nucleic acid molecules comprising truncated forms of the human cytomegalovirus (CMV) promoter are operably linked to a transgene of interest, including those encoding eNOS protein are taught. Vectors comprising these nucleic acid molecules and host cells transformed by these vectors and methods of producing cellular compositions are used for the treatment of a variety of pulmonary and cardiac diseases. There is provided a truncated human cytomegalovirus (CMV) enhancer element comprising SEQ ID NO: 1 or a functional derivative thereof. A truncated human cytomegalovirus (CMV) promoter comprising the truncated CMV enhancer element is taught. A polynucleotide expression cassette comprising the truncated CMV promoter and a transcribable polynucleotide operably linked to the truncated CMV promoter polynucleotide construct, a host cell comprising the polynucleotide expression cassette, and use of the host cell treat renal, vascular, pulmonary, or cardiac disease in the patient are taught.

This invention discloses a recombinant vector containing the LjARF25 gene from honeysuckle and its applications, belonging to the field of plant genetic engineering technology. The recombinant vector is based on the pCAMBIA series of binary plant expression vectors, containing an endogenous auxin-responsive promoter 2000 bp upstream of the LjARF25 gene as shown in SEQ ID NO:2 and the coding region sequence of the LjARF25 gene as shown in SEQ ID NO:1, constructing an LjARF25pro::LjARF25 inducible expression cassette. The LjARF25 gene responds to exogenous auxin stimulation, and its expression level shows significant time- and concentration-dependent changes after exogenous auxin treatment. This invention, by constructing an LjARF25 gene inducible expression vector suitable for honeysuckle, provides a technical foundation for honeysuckle gene function research, metabolic regulation of medicinal components, and molecular breeding.

This invention discloses a method for enhancing salt tolerance in rice by knocking out the OsAKIP30 gene, belonging to the field of plant genetic engineering technology. This invention utilizes CRISPR / Cas9 technology to target and mutate this gene. Specific steps include: selecting target sequences T1 and / or T2 in the CDS region of the OsAKIP30 gene; constructing a recombinant vector pYLCRISPR / Cas9Pubi-B-OsAKIP30 containing the corresponding sgRNA and Cas9 expression cassette; transforming rice varieties using Agrobacterium-mediated transformation; identifying mutant plants through genomic PCR and sequencing; and identifying the salt tolerance phenotype of the mutants (e.g., seedling survival rate after salt treatment, ion leakage rate, and chlorophyll content), screening for lines with superior salt tolerance compared to the wild type. This invention provides an effective technical approach for salt-tolerant rice breeding.

Herein is reported a method for producing recombinant adeno-associated viral particle preparation (rAAVp) comprising the step of cultivating a mammalian cell comprising expression cassettes for a non-adeno-associated viral gene, which is interspaced between two AAV inverted terminal repeats (ITRs), an adeno-associated virus rep gene, an adeno-associated virus cap gene, an adeno-associated virus E1A gene, an adeno-associated virus E1B gene, an adeno-associated virus E2A gene, an adeno-associated virus E4orf6 and an adeno-associated virus VA RNA gene, and thereby producing the rAAVp, wherein the cultivating is at a pH value in the range of and including pH 7.4 to pH 7.6. The yield of the rAAVp produced by the cultivating at a pH value in the range of and including pH 7.4 to pH 7.6 is higher than the yield of a rAAVp produced by a cultivating at a pH value in the range of and including pH 7.0 to pH 7.2 and the rAAVp produced by the cultivating at a pH value in the range of and including pH 7.4 to pH 7.6 has a higher percentage of full particles than a rAAVp produced by a cultivating at a pH value in the range of and including pH 7.0 to pH 7.2.

The application discloses application of ZmHB77 protein and a coding gene thereof in regulation of drought resistance of plants. The application discloses application of ZmHB77 protein (SEQ ID No. 1) or a related biological material thereof in all or part of the following: regulation of drought resistance of plants, regulation of seed root number of plants, regulation of lateral root density of plants, and the related biological material is a nucleic acid molecule capable of expressing the ZmHB77 protein or an expression cassette containing the nucleic acid molecule, a recombinant vector, a recombinant bacterium or a transgenic cell line. After gene knockout of the ZmHB77 gene drought resistance protein coding gene of the application, the drought resistance of corn is significantly improved. The application provides a more precise, efficient and safe technical method for creating a corn overexpression plant and / or corn breeding, realizes precise improvement of the drought resistance of corn, can promote the commercial corn breeding process, overcomes the short board of traditional breeding, shortens the breeding cycle and does not affect other properties.