40 results about "GENE RE-ARRANGEMENTS" patented technology

The invention relates to improved methods for directed evolution of polymers, including directed evolution of nucleic acids and proteins. Specifically, the methods of the invention include analytical methods for identifying "crossover locations" in a polymer. Crossovers at these locations are less likely to disrupt desirable properties of the protein, such as stability or functionality. The invention further provides improved methods for directed evolution wherein the polymer is selectively recombined at the identified "crossover locations". Crossover disruption profiles can be used to identify preferred crossover locations. Structural domains of a biopolymer can also be identified and analyzed, and domains can be organized into schema. Schema disruption profiles can be calculated, for example based on conformational energy or interatomic distances, and these can be used to identify preferred or candidate crossover locations. Computer systems for implementing analytical methods of the invention are also provided.

The invention discloses gene recombination mediated on the basis of phiC312 and inheritance reformation for an erythrocin producing bacterium. A target gene can be stably integrated on a locus pre-constructed by a host cell genome by a method of gene recombination mediated by a phiC312 locus specificity integration mechanism. The invention also relates to an application of the method on the inheritance reformation of the erythrocin producing bacterium.

The invention discloses gene-recombinant saccharomyces cerevisiae for efficiently adsorbing uranium contained in a water solution and a construction method thereof. The gene-recombinant saccharomyces cerevisiae is constructed by carrying out codon reconstruction on a human liver metallothionein gene, recombining with a saccharomyces cerevisiae expression vector and transferring into a saccharomyces cerevisiae cell for correct expression. The gene-recombinant saccharomyces cerevisiae disclosed by the invention effectively increases the adsorption capacity of the saccharomyces cerevisiae cell on uranium (VI), has the characteristics of low adsorption cost and biomass reusability without secondary pollution and has positive promoting effect on the aspects of uranium pollution degree reduction and uranium resource recovery.

The invention discloses a power marketing inspection method and a power marketing inspection system based on gene recombination and feature clustering. The method comprises the following steps: 100, data cleaning: carrying out the provincial inspection of imported work order data, and generating two results after the first data cleaning, i.e., an effective abnormality and an invalid abnormality ofthe work order data; 200, issuing the valid and abnormal data to a municipal unit for data processing, and directly processing the invalid and abnormal data; and 300, carrying out intelligent processing on the data of the municipal units through an intelligent inspection method. According to the invention, the accuracy of on-line inspection can be effectively improved, and the efficiency of on-line inspection operation is greatly improved.

The invention discloses a vibrio mimicus efficient genetic combination method based on natural conversion and application. The method comprises the steps that a vibrio mimicus SCCF01 plant genome is adopted as a template, a target gene upper arm and target gene lower arm primer pair is adopted for amplifying a gene upper arm sequence and a lower arm sequence; a kanamycin resistance gene segment is amplified; the kanamycin resistance gene segment obtained through amplification, the target gene upper arm sequence and the lower arm sequence segments are fused, and a fusion PCR product containing the kanamycin resistance gene segment, the target gene upper arm sequence segments is obtained; the obtained PCR product and the acceptor vibrio mimicus SCCF01 plant are subjected to natural conversion, and the vibrio mimicus SCCF01 plant deleted plant is obtained by screening a resistance marker gene. Compared with the suicide plasmid mediated gene recombination technology and the lambda-RED recombination technology, no electricity conversion plasmid or targeting DNA is needed in the operation process, and operation is easy.

The invention relates to a production method of a gene recombinant protein Tat-hMsrA. The production method comprises the following steps that recombinant plasmid containing a fusion protein Trx-Tat-hMsrA is constructed; the recombinant plasmid is electrically transformed into a host, namely bacillus subtilis for strain culturing to obtain a positive expression fungus; and the positive expressionfungus is subjected to induction culture, expression product liquid containing Tat-hMsrA is obtained, digestion, renaturation and purification are conducted, thus the gene recombinant protein Tat-hMsrA is obtained. According to the production method of the gene recombinant protein Tat-hMsrA, the Tat-hMsrA with high activity can be obtained.

The invention provides a method for constructing and producing recombination III-type restriction endonuclease Ecop15I. The III-type restriction endonuclease Ecop15I plays an important role in serial analysis of gene expression, and the prior method is difficult to realize large-scale expression and purification. By utilizing a gene recombination method, two subunit fragments of the Ecop15I fused with a purification tag are artificially synthesized and are cloned into two independent expression units of a prokaryotic co-expression vector pACYCDuet-1 respectively; under IPTG induction, the expression units with independent T7 promoters, ribosome binding sites, start codon and stop codon simultaneously and independently express two subunits of enzyme in Escherichia coli, and are folded into a composite structure with biological activity; purification is performed through Ni affinity chromatography and DEAE ion-exchange chromatography; and the recombination-type Ecop15I restriction endonuclease with high yield and good activity and stability is finally obtained. Therefore, the method provides an effective way for mass production.

The invention discloses a building method and application of a lactobacillus reuteri resistance-marker-free gene integration system. By means of the point mutation technology, a point mutation gene pheSM of a lactobacillus reuteri pheS gene is obtained, the point mutation gene pheSM and a erythromycin resistance gene are used for building an integrated framework LR-pheSM-em-RR, the integrated framework LR-pheSM-em-RR is electrically converted into L.renteri XNY competent cells, and pheSM genetic recombination lactobacillus L.renteri XNYM is obtained through erythromycin screening; then, the gene integration framework is constructed and inserted, then the gene integration framework is electrically converted into pheSM gene recombination lactobacillus reuteri competent cells, and the lactobacillus reuteri without resistance markers can be obtained through para-chlorophenylalanine acid resistance screening. According to the method, resistance genes are not introduced into host bacteria genomes, so that biological safety hidden hazards of the resistance genes and the polarity effect of the resistance genes on upstream genes and downstream genes are avoided. By means of the system, gene targeted integration can be carried out on the host bacteria for multiple times.

The invention provides a preparation method of a recombination biological film for promoting gene editing T cell activation and amplification. The recombination biological film is prepared through 1),leading a modified CD137L gene sequence to a K562 cell unit through a gene recombination method to obtain a steady transfer strain cell of CD137L protein stably expressed and modified at the K562 cell surface, wherein the cell is named as rCD137L-K562 cell; 2), regularly culturing the rCD137L-K562 cell in step 1), and breaking the cell through the cell lysis buffer; carrying out a differential speed centrifugal method to obtain the biological film containing the modified CD137L protein; gathering, extracting and purifying the biological film containing the modified CD137L protein by an affinity chromatography and a magnetic bead separating method. The preparation method can improve the irritation of the gene editing T cell, and is free from any bad and side effects.

The invention relates to a T1R3 gene overexpressed lentiviral vector and a T1R3 lentivirus, and construction methods thereof, and belongs to the technical field of molecular biology. By a gene recombination technology, construction is performed by taking a pLVX-Puro lentiviral expression vector as the basis, a T1R3 gene is integrated, and the T1R3 gene overexpressed lentiviral vector pLVX-Puro-T1R3 is obtained. Lentiviral packaging plasmids pRSV-Rev, pMDlg-pRRE and pMD2.G are used for packaging the T1R3 gene overexpressed lentiviral vector pLVX-Puro-T1R3, and HEK293 cells are transfected to obtain the T1R3 lentivirus. The constructed T1R3 gene overexpressed lentiviral vector has high transfection efficiency to host cells, and can specifically, continuously and efficiently promote stable expression of the T1R3 gene in the host cells.

The invention provides an interface-controllable material gene recombination blending modification technology. The technology mainly comprises a material gene recombination multi-unit plasticizing extrusion device, a stretching device, a double-screw blending device and a granulation device. The technology is based on a material gene recombination principle, various monomer materials are combined and matched in advance according to given relative positions and proportions, and the preparation technology is combined, so that the proportion and the interface state of the multi-component materials are controlled, and the gene recombination composite material with specific performance is obtained. According to the specific implementation mode, firstly, monomer materials are combined and co-extruded according to the given relative position and proportion through a material gene recombination multi-unit plasticizing extrusion device, after extrusion, the composite material of the multi-layer structure is obtained, the interface layers of the monomer materials of each layer are distinct, then the composite material structure is stretched and refined through a differential roller in a stretching device, and finally the composite material is obtained. Then, a composite material with specific performance is obtained under the plasticizing action of a double-screw blending device, and finally, granules are obtained through a granulating device.

The invention provides an m6A'reader 'YTHDF2 gene modification method and application thereof, and relates to the technical field of gene engineering. According to the gene modification method, a gene editing technology is used for modifying a YTHDF2 gene to obtain a cell line with the YTHDF2 gene knocked out, and the gene modification method comprises the steps of sgRNA sequence design, YTHDF2 gene knockout, plasmid construction and modified gene recombination cell line screening. According to the invention, the cells with specific genes are subjected to gene modification, the fact that the YTHDF2 gene is knocked out, inserted and subjected to base mutation by using the CRISPR/Cas9 technology in the Vero81 cells is proposed for the first time, a brand new concept is provided for cytology or genetic research, and the application prospect is very wide; and according to the Vero cell line YTHDF2 gene modification method provided by the invention, the mRNA methylation level in host cells can be changed, so that the virus multiplication capacity is changed, and the possibility is provided for researching the relationship between the virus and host methylation.

The invention provides a gene recombination method mediated by VWB-attP and [phi]C31-attP integrase and an application thereof. Specifically, the two integrase modules are constructed in the same plasmid containing a target gene by utilizing a site-specific integration mechanism mediated by a VWB-attP integrase system and a [phi]C31-attP integrase system; and a target gene needing to be expressed can be rapidly recombined to a corresponding attB site of a streptomyces chromosome in a multi-copy number through inter-genus conjugation transfer. The method disclosed by the invention is applied to abamectin, erythromycin and lincomycin producing bacteria.

Production of gene-recombinant fused protein K1-EN is carried out by constructing for recombinant plasmid pPICZ alpha A-K1-EN, constructing for recombinant yeast gene engineering bacterial pPICZ alpha A-K1-EN/GS115, expressing for recombinant fused protein K1-EN and purifying recombinant fused protein K1-EN. Pichia is eukaryotic expression system with methanol as the only carbon resource; pPICZ alpha A contains Paox1, which is secretory expression plasmid and expresses accidental protein under methanol induction. It is cheap and no need for externally heating source and has large scale.

The invention relates to a gene recombinant plasmid, gene recombinant pichia pastoris and straw fiber degumming application, and belongs to the technical field of renewable resource utilization. The gene recombinant plasmid comprises a constitutive promoter, a ribosome binding site, a pectin degrading enzyme gene modified by a first signal peptide gene, a terminator, a first acid-sensitive promoter, a ribosome binding site, a small laccase gene modified by a second signal peptide gene, a terminator and a first acid-sensitive promoter which are sequentially connected from upstream to downstream, the kit comprises a first acid-sensitive promoter, a ribosome binding site, a peroxidase gene, a terminator, a second acid-sensitive promoter, a basic gene and a terminator. According to the method, pectin and cellulose in the straw are degraded by using the recombinant pichia pastoris, the comprehensive utilization rate of the straw is improved, high-quality and zero-pollution straw fibers for spinning and derivative chemical products are produced, and the yield of industrial products with high additional value can be increased while the waste straw treatment cost and labor force can be reduced.

The invention belongs to the technical field of gene recombination and protein expression, and particularly relates to an African green monkey kidney cell line capable of stably expressing SLAM protein as well as a construction method and application of the African green monkey kidney cell line, the cell line is African green monkey kidney cells Vero-E6-SLAM, the African green monkey kidney cells Vero-E6-SLAM are sent to Guangdong Microbial Culture Collection Center on November 11, 2021 to be preserved, and the preservation number is GDMCCNO: 62054. The preparation method comprises the following steps: constructing a pAAVS1-Donor-SLAM overexpression plasmid, and then co-transfecting the pAAVS1-Donor-SLAM overexpression plasmid and pCass-Guide-AAVS1 into a Vero-E6 cell, so as to obtain the recombinant vector. According to the cell line, an exogenous gene can be integrated into a known safe site in a genome, normal expression of the transferred gene is realized without influencing the normal physiological function of the cell, the cloning accuracy is high, the cell line can be applied to high-yield SLAM protein, and a platform is provided for separation and pathogenic mechanism research of canine distemper virus.

In immunoassay using a latex reagent, the correctness of immunoassay in a CRP high-concentration range is improved. The recombinant C-reactive protein is a C-reactive protein produced by gene recombination, and 55% or more of the N-terminus of the C-reactive protein is pyroglutaminated.

The invention discloses gene recombinant MSCs (mesenchymal stem cells) with a function of promoting hair regeneration and a preparation method of an exosome-like nano material of the gene recombinant MSCs. The preparation method comprises the steps: S101, preparing umbilical cord mesenchymal stem cells (hU-MSCs); S102, constructing a Jag1 membrane surface display plasmid (pDJag1); S103, constructing a miR-218-5p miRNA overexpression plasmid; S104, expressing Jagged 1 proteins on the surfaces of cell membranes, and expressing gene recombination MSC (JagmirMSC) cells of miR-218-5p miRNA in the cells; and S105, further processing the JagmirMSC cells into the exosome-like nano material. According to the invention, the JagmirMSC cells and the exosome-like nano material thereof have the function of promoting hair regeneration. The JagmirMSC cells exosome-like nano material not only has the same function of promoting hair regeneration as the JagmirMSC cells, but also is convenient to store, transport and use, is also suitable for mass production, and has a greater industrialization potential.

The invention discloses a preparation method of gene recombinant collagen oligopeptide. The preparation method of the gene recombinant collagen oligopeptide comprises the following steps: S1, synthesizing an MYS-1 gene by adopting a pair of complementary primers through PCR (Polymerase Chain Reaction); s2, carrying out double enzyme digestion on the MYS-1 gene to obtain a recombinant plasmid; s3, transforming expression host escherichia coli by using the recombinant plasmid to obtain expression engineering bacteria; s4, inducing the expression engineering bacteria to express fusion protein; s5, performing purification: collecting an eluent; s6, enzyme digestion of the fusion protein and purification of a target polypeptide MYS-1; and S7, purifying and preparing the collagen oligopeptide MYS-1 by using a high performance liquid chromatography technology to obtain the high-purity gene recombinant high-stability collagen oligopeptide MYS-1. The length of the prepared collagen oligopeptide is far smaller than that of natural human collagen and related hydrolyzed polypeptide, the collagen oligopeptide is small in molecular weight, easy to prepare and high in efficiency, and the collagen oligopeptide has good biological activities such as oxidation resistance, aging resistance, repair promotion and melanogenesis inhibition.