12009results about "Nucleic acid vector" patented technology

Disclosed herein are modified split mirrored transposons or double-stranded split mirrored transposon (SMT) compositions and methods of using SMTs. The methods include producing SMTs and using SMTs for nucleic acid analysis.

Methods and systems for sample preparation techniques that prevent inhibition of reactions such as due to the presence of longer oligonucleotides are presented herein. The methods and systems provided herein allow amplification (e.g., whole genome amplification) and sequencing of chromatin accessible regions of single cells.

The invention belongs to the technical field of bioengineering, and discloses a recombinant yarrowia lipolytica capable of efficiently and rapidly producing sclareol, which is characterized in that optimized sclareol synthase related coding genes ScLPPS, ScSCS, GGPPS1, GGPPS2, ERG20 and tHMGR coding genes are inserted into a yarrowia lipolytica genome; inserting optimized pentadienol utilization pathway coding genes such as IPK, THIM and IDI coding genes; and inserting an optimized ATP: citrate lyase coding gene ACLY to obtain the ATP: citrate lyase coding gene ACLY. The recombinant yarrowia lipolytica is based on knockout of a yarrowia lipolytica strain responsible for coding a non-homologous recombination gene ku70, so that the homologous recombination capability of the yarrowia lipolytica strain is enhanced, gene integration is realized through the homologous recombination function of the yarrowia lipolytica, the genetic stability of the introduced gene can be greatly improved, and the recombinant yarrowia lipolytica strain has a good application prospect. The construction method of the recombinant yarrowia lipolytica strain is efficient and simple to operate.

To provide methods for treating a subject having or at risk of developing a neurocognitive disorder, such as frontotemporal lobar degeneration or neuronal ceroid lipofuscinosis.SOLUTION: Provided is a method comprising administering, to a subject diagnosed as having a neurocognitive disorder (NCD), a composition comprising a population of cells comprising a transgene encoding a progranulin (PGRN) or a granulin (GRN).SELECTED DRAWING: Figure 1A

To provide a medicine for treating a disease in a mammal.SOLUTION: Provided is a medicine for treating a disease in a mammal, comprising an expression cassette that is a first expression cassette comprising a chimeric gene operably linked to a first expression control element, the chimeric gene comprising a first portion comprising a minigene having an alternatively spliced exon and a second portion encoding a first RNA encoding a protein, the first portion being 5' to the second portion, the second portion lacking an initiation codon or start codon, the alternatively spliced exon comprising a translation initiation regulatory sequence, expression of the protein being controlled by inclusion of the alternatively spliced exon, and the medicine being used in combination with a small molecule splicing modifier that modulates splicing of the alternatively spliced exon.SELECTED DRAWING: Figure 1

A recombinant humanized collagen and a preparation method therefor. The recombinant humanized type III collagen can be efficiently and stably expressed in three expression systems of yeast, Escherichia coli, and CHO cells, and has the biological activity superior to that of a commercially available collagen product. The recombinant humanized type III collagen has structural characteristics and functional activity similar to those of a natural collagen, and thus has wide application prospects in biomedical materials, tissue engineering products, cosmetics, foods, health care products or drugs, medical instruments, medical cosmetology and the like.

The invention belongs to the technical field of optimized coding genes, and discloses a recombinant human I-type collagen as well as a preparation method and application thereof, and the amino acid sequence of the recombinant human I-type collagen is shown as SEQ ID No.1. The recombinant I-type collagen provided by the invention is relatively high in expression quantity, has relatively good cell migration promoting, cell proliferation promoting and cell adhesion promoting properties, is high in safety, and can be widely applied to the fields of medicines, medical instruments, biological materials, tissue engineering, cosmetics and the like.

The invention discloses a bovine serum albumin combined nano antibody and a preparation method and application thereof, the amino acid sequence of the bovine serum albumin combined nano antibody Nb1 is shown as SEQ.ID No.1, the nano antibody Nb not only has high affinity to bovine serum albumin, but also has good binding activity to both human serum albumin and mouse serum albumin, and the nano antibody Nb has high affinity to human serum albumin and mouse serum albumin. And the antibody has a relatively good broad-spectrum application range, and can be widely applied to the aspects of drug half-life period enhancement, tumor targeted therapy and imaging, immunodetection or diagnosis, recombinant albumin purification or enrichment and the like.

The invention provides recombinant human III-type collagen which is of a triple-helix structure and assembled into collagenous fibers and application of the recombinant human III-type collagen, and belongs to the technical field of bioengineering. Collagen fragments capable of forming a triple helix structure are screened out through a bioinformatics algorithm, the fragments are connected in series, a key C-terminal structural domain and a specific amino acid sequence are combined, and formation of a stable triple helix advanced structure and assembly of collagenous fibers are effectively promoted. The recombinant human III type collagen is efficiently expressed through a eukaryotic expression system, and shows excellent stability, low immunogenicity and good biocompatibility. Experimental results show that the prepared recombinant human III-type collagen not only has a natural triple helix structure, but also can be further assembled into macromolecular collagen fibers, is suitable for multiple fields such as medical beauty, bioengineering and tissue repair, and has wide application prospects and market values.

Engineered CRISPR-Cas9 nucleases with altered and improved PAM specificities and their use in genomic engineering, epigenomic engineering, and genome targeting.

The invention discloses a recombinant humanized XVII type collagen as well as an expression vector and a genetically engineered bacterium thereof. Four functional fragments with high activity are obtained through screening, one of the fragments is optimized and then combined according to a certain sequence to obtain recombinant humanized XVII type collagen JYC1703, or one of the fragments is optimized and then combined according to a certain sequence, and GPP sequences are fused at an amino terminal and a carboxyl terminal respectively to obtain recombinant humanized XVII type collagen JYC1701 and JYC1702, and the recombinant humanized XVII type collagen JYC1701 and JYC1702 are combined according to a certain sequence to obtain the recombinant humanized XVII type collagen JYC1701 and JYC1702. After codon optimization, amplifying and cloning into an expression vector, transforming into pichia pastoris competent cells after enzyme digestion linearization, and screening high-copy recombinants through G418 resistance gradient, so as to obtain pichia pastoris genetically engineered bacteria for expressing recombinant humanized XVII type collagen. The recombinant humanized XVII type collagen JYC1701, JYC1702 and JYC1703 disclosed by the invention show excellent functions of promoting cell adhesion, cell proliferation and cell migration, and are suitable for preparing products such as skin care products or medical instruments and the like.

The invention discloses a recombinant herpes zoster vaccine as well as a preparation method and application thereof, and particularly relates to the technical field of biological medicines. The recombinant herpes zoster vaccine comprises a recombinant herpes zoster vaccine fusion protein purified product HN101 and a liposome mixture HA201, and the recombinant herpes zoster fusion protein gE-gD is constructed by recombining reconstructed gE protein and reconstructed gD protein. The gE-gD recombinant herpes zoster fusion protein is obtained by reconstructing gE protein of varicella-herpes zoster virus and gD protein of HSV through a gene recombination technology and then recombining the gE protein and the gD protein. After the gE-gD recombinant herpes zoster fusion protein is combined with a liposome mixture HA201, stronger immune response can be induced, the antibody titer and cellular immunity are improved, the vaccine effect is enhanced, and meanwhile, the potential infection risk is avoided.

The present invention relates to a method for the production and purification of a protein of interest, and more particularly, the present invention relates to a fusion protein comprising: a self-aggregating peptide, a linker peptide comprising an amino acid sequence selected from an elastin-like polypeptide (ELP) or a protease propeptide (ProP) and the spacer comprising a cleavage site, and a protein portion of interest, and a method for column-free one-step purification of a target protein by expressing the fusion protein.

The invention discloses a fusion enzyme capable of efficiently degrading PET (polyethylene terephthalate) plastic and application of the fusion enzyme. The invention provides a recombinant fusion enzyme. The recombinant fusion enzyme comprises a mutant KL-MHETase of thermophilic carboxylesterase Est30 and a PET (Polyethylene Terephthalate) degrading enzyme FAST-PETase, the amino acid sequence of the mutant KL-MHETase of the thermophilic carboxylesterase Est30 is as shown in SEQ ID NO. 3; the amino acid sequence of the FAST-PETase is as shown in the 284th to 546th amino acids of SEQ ID NO.1 (Sequence Identifier Number 1). The fusion enzyme is used as a biocatalyst, the activity of catalyzing hydrolysis of MHET at 50 DEG C is 52.6 times that of FAST-PETase, and the fusion enzyme has high enzyme activity and thermal stability. PET degradation is catalyzed at the temperature of 50 DEG C, the conversion rate of PET is 90% after 48 hours of reaction, and meanwhile, the TPA content in the product is close to 100%.

The invention relates to an application of oxygenase in catalyzing 4-ethene guaiacol to synthesize vanillin. The amino acid sequence of the oxygenase comprises a sequence as shown in SEQ ID NO.1. The invention also relates to an application of the oxygenase in catalyzing 4-ethene guaiacol to synthesize vanillin. The invention provides an application of oxygenase in catalyzing synthesis of vanillin from 4-ethene guaiacol. The 4-ethene guaiacol oxygenase is in soluble expression in host bacteria. Crude enzyme liquid expressed by the strain stably catalyzes conversion of 4-ethylene guaiacol into vanillin, the conversion rate within 2 hours is 80%, the conversion rate within 24 hours exceeds 98%, and 11.5 g / L of vanillin can be accumulatively produced within 6 hours.

The invention belongs to the technical field of biology, and provides a porcine epidemic diarrhea virus mRNA vaccine and application. The mRNA molecule comprises a coding region for coding the S protein of the PEDV; the PEDV is of a G2a type, and two proline mutations I1076P and L1077P are introduced into an S protein. The invention further discloses a DNA molecule. An S protein of PEDV; a recombinant plasmid; recombining the host cell; an mRNA vaccine formulation which can be used to prepare an antibody against the S protein of PEDV after immunizing an animal; the invention further discloses application of the mRNA molecule, the DNA molecule, the recombinant plasmid or the host cell to preparation of a medicine for preventing PEDV (Porcine Epidemic Diarrhea Virus) infection. The vaccine prepared by the invention is fast in antibody generation and high in protection rate, can generate high-level neutralizing antibodies in serum and breast milk of sows after the pregnant sows are immunized before parturition, has a remarkable protection effect on piglets, and can effectively prevent the piglets from being infected with PEDV (Porcine Epidemic Diarrhea Virus).

The invention relates to retrieving or modifying target nucleic acids, such as host cell chromosomal DNA, by homologous recombination with vectors that have been cut to provide recombinogenic nucleic acid strands in situ. The methods described herein can be used to modify a target nucleic acid in vitro or in any prokaryotic or eukaryotic cell using homologous recombination.

The invention relates to a saccharomyces cerevisiae strain for efficiently synthesizing cordycepin as well as a construction method and application of the saccharomyces cerevisiae strain, and belongs to the technical field of microbial genetic engineering. The construction method provided by the invention comprises the following steps: carrying out multi-copy genome integrated expression on heterologous codon optimized 2 '-carbonyl-3'-deoxyadenosine reductase genes and 3 '-adenosine monophosphate phosphohydrolase genes, so as to increase the dosage of key enzymes; and supplementing the defective gene of the engineering strain to obtain the saccharomyces cerevisiae strain B5U3 with high yield of cordycepin. According to the obtained saccharomyces cerevisiae strain for efficiently synthesizing the cordycepin, the cordycepin yield reaches 2.92 g / L after shake flask fermentation, the cordycepin yield reaches 7.5 g / L after fermentation in a fermentation tank, and a foundation is laid for safer and more reliable industrial production of the cordycepin.

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.

Provided herein are gene editing pairs comprising an engineered protein, e.g, an engineered CRISPR protein and a guide ribonucleic acid (gRNA) that may provide improved gene editing relative to reference, i.e., wild type sequences. In some embodiments, the engineered protein of the gene editing pair comprises a RuvC cleavage domain with one or more amino acid modifications relative to a reference CRISPR protein RuvC sequence. In some embodiments, a guide ribonucleic acid (gRNA) of the gene editing pair comprises at least one modification in a scaffold stem loop relative to a reference scaffold stem loop sequence.

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 belongs to the field of agricultural biotechnology and environmental microbiological engineering, and discloses mesotrione nitroreductase Mstnr as well as a coding gene and application thereof. The nucleotide sequence of the gene is SEQ ID NO.1, the full length is 678bp, 225 amino acids are encoded, and the amino acid sequence of the gene is SEQ ID NO.2. The highest homology of the Mtnr and reported nitroreductase is only 49.2%, so that the Mtnr is a new nitroreductase gene. The nitroreductase Mtnr provided by the invention can catalyze nitro reduction of mesotrione and convert mesotrione into a product 2-amino-4-methylsulfonylbenzoic acid (AMBA) without herbicidal activity, which indicates that the Mtnr gene has a huge application prospect in biodegradation detoxification and repair of mesotrione.

The invention discloses a high-activity and high-thermal-stability MHET hydrolase mutant and application thereof. The invention provides a mutant protein of thermophilic carboxylesterase, which is shown as 1) or 2): 1) the protein shown as 1) is a protein obtained by mutating amino acid residues at the 130th site and the 171th site of wild thermophilic carboxylesterase shown as SEQ ID NO.2 or a sequence corresponding to the SEQ ID NO.2 and keeping other amino acid residues unchanged; (2) the protein is obtained by mutating amino acid residues at the 127th site, the 130th site and the 171th site of wild thermophilic carboxylesterase and keeping other amino acid residues unchanged. Experiments prove that the activity of the modified mutant for catalyzing MHET hydrolysis at the temperature of 50 DEG C is improved by 95 times compared with that of a wild type, and the modified mutant has higher enzyme activity and thermal stability and is the first enzyme capable of efficiently catalyzing MHET hydrolysis at the temperature of 50 DEG C at present.

The present invention provides a method for packaging a BaEV retroviral vector. The method includes constructing a stable BaEV retroviral packaging cell line, transfecting a retroviral vector plasmid containing a target sequence into a virus-producing cell line, where the virus-producing cell line is a HEK293T cell line or its derivative cell line, to harvest the transiently transfected and virus-producing retroviral vector, mixing it with the stable BaEV retroviral packaging cell line, and promoting the transduction of the retroviral vector into the stable BaEV retroviral packaging cell line by horizontal centrifugation. The present invention can achieve the packaging of a BAEV-type retroviral vector carrying a CD19CAR foreign gene plasmid, construct a BAEV-type retroviral vector-producing cell line, and produce a high-titer BAEV-type retroviral vector.

The invention belongs to the technical field of bioengineering, and particularly relates to recombinant III-type humanized collagen as well as a coding gene and application thereof. According to the invention, a novel III-type humanized collagen sequence is designed by adopting an advanced AI technology and molecular dynamics simulation, so that the recombinant III-type humanized collagen product is ensured to have high stability and degradation resistance. According to the invention, the screened new III-type humanized collagen sequence is transferred to the pichia pastoris strain for recombinant expression, and 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.

The invention provides a high-stability recombinant humanized elastin, a genetically engineered bacterium for expressing the protein and application of the high-stability recombinant humanized elastin, and belongs to the technical field of binding tissue peptides. The nucleotide sequence of the recombinant humanized elastin is shown as SEQ ID NO.1; and the amino acid sequence is as shown in SEQ ID NO.4. According to the invention, the nucleotide sequence of the humanized elastin is subjected to codon optimization and transferred into escherichia coli to construct the genetically engineered bacterium for expressing the recombinant humanized elastin, so that the expression quantity of the recombinant humanized elastin can be increased, the stability of the recombinant humanized elastin can be improved, and the recombinant humanized elastin can be used for preparing the recombinant humanized elastin. The migration rate of the recombinant humanized elastin to the HSF cell is improved, and the expression quantity of the h-ELN gene, the h-COL1A1 gene and the h-COL3A1 gene of the HSF cell is improved.

The invention discloses an anti-interleukin-6 antibody or a reagent and a kit for detecting interleukin-6, and relates to the field of antibodies. The anti-interleukin-6 antibody disclosed by the invention comprises a heavy chain complementarity determining region and a light chain complementarity determining region, provides an important raw material source for detection of interleukin-6, and has good affinity or activity.

The invention provides a porcine epidemic diarrhea virus S protein mutant, a nucleic acid molecule, a subunit vaccine and application. Compared with wild type porcine epidemic diarrhea virus S protein, the porcine epidemic diarrhea virus S protein mutant has the advantage that amino acid at the 1076th site, amino acid at the 1077th site, amino acid at the 1016th site and amino acid at the 1017th site are mutated into proline. The natural trimer which is stable and maintains the conformation of the S protein before fusion is constructed, the safe and efficient PEDV subunit vaccine is obtained, and the use effect and immunogenicity of the porcine epidemic diarrhea vaccine can be improved; moreover, compared with a wild type S protein, the porcine epidemic diarrhea virus S protein mutant disclosed by the invention has a higher expression quantity.