115 results about "Cytoplast" patented technology

The invention belongs to the technical field of genetic breeding, and relates to an InDel marker for identifying green Chinese onion cytoplasm fertility based on a chloroplast genome and application of the InDel marker. The InDel marker is subjected to amplification detection by adopting a primer group, and the primer group comprises one or more of a first primer group, a second primer group, a third primer group and a fifth primer group; a pair of nucleotide sequences of the first primer group are as shown in SEQ ID NO.1-2; a pair of nucleotide sequences of the second primer group are as shown in SEQ ID NO.3-4; a pair of nucleotide sequences of the third primer group are as shown in SEQ ID NO.5-6; a pair of nucleotide sequences of the fourth primer group are as shown in SEQ ID NO.7-8; and a pair of nucleotide sequences of the fifth primer group are as shown in SEQ ID NO.9 to SEQ ID NO.10. The InDel marker provided by the invention can be used for identifying the cytoplasm fertility of the green Chinese onion, and the tedious procedure and process for extracting DNA can be omitted in the detection process, so that the detection process is simplified.

The invention discloses an HPV whole genome stable strain as well as a preparation method and application thereof, and relates to the field of gene recombination. The invention provides a method for constructing an HPV (human papillomavirus) whole genome stable strain by utilizing a PiggyBac transposon system. The sequence of an HPV whole genome is shown as SEQ ID NO. 2. The HPV whole genome stable strain obtained by the method has high stability and high copy number, and can be used as an HPV genome cell quality control product.

The invention discloses a method for losing high-temperature rejuvenation phenotypes of wild abortion type cytoplasmic male sterile line rice sensitive to high temperature. The method comprises the following steps: replacing a DNA fragment Rf3-TOS in the wild abortion type cytoplasmic male sterile line rice sensitive to high temperature into a DNA fragment Rf3-nonTOS, so that the high-temperature rejuvenation phenotype of the wild abortion type cytoplasmic male sterile line rice sensitive to high temperature is lost; the nucleotide sequence of the DNA fragment Rf3-TOS is shown as SEQ ID No. 1 in a sequence table; the nucleotide sequence of the DNA fragment Rf3-nonTOS is as shown in SEQ ID No.2 in a sequence table; the wild abortion type cytoplasmic male sterile line rice sensitive to high temperature is wild abortion type cytoplasmic male sterile line rice with a high-temperature restoration phenotype. The invention lays a solid foundation for the safety of hybrid rice seed production.

The invention relates to the technical field of biology, in particular to a chicken CKAP4 protein polyclonal antibody and application and a kit thereof. An animal is immunized with the CKAP4 recombinant protein, serum is purified from the immunized animal, the chicken CKAP4 protein polyclonal antibody is obtained, and the amino acid sequence of the CKAP4 recombinant protein is shown as SEQ ID No.1. Sensitivity and specificity detection tests on the chicken CKAP4 protein polyclonal antibody find that the serum titer of CKAP4 is 1: 102400, the expression of the CKAP4 protein mainly focuses on cytoplasm parts of various organs of animals, and the polyclonal antibody can specifically bind to the CKAP4 protein in animal tissues. Therefore, the polyclonal antibody has the advantages of high sensitivity, good specificity, wide application range and the like, and a foundation is laid for later detection of CKAP4 protein and research on pathological mechanisms of virus infection diseases.

Heterologous synthesis of a nitrogen-fixing system in a non-diazotrophic organism is a long-sought-after goal because of the significance of nitrogenase for areas related to agronomy, energy and environment. Two simplified nitrogenase analogs from Azotobacter vinelandii in Escherichia coli were successfully heterologously synthesized. One analog comprise the reductase component (NifH) and, in place of the catalytic component (NifDK), a cofactor maturase (NifEN); whereas the other analog comprised only NifEN. Metal, EPR and activity analyses verified the cluster composition and functional competence of the heterologously expressed NifH and NifEN. Moreover, NMR, nanoSIMS and growth experiments illustrated the abilities of both systems to mimic the nitrogenase in reducing N2 and incorporating the reduced N into the cellular mass. These results firmly established NifEN / NifH or NifEN as a simplified nitrogenase analog that could be engineered or expanded to facilitate future transgenic expression of nitrogenase and development of nitrogenase-based biotechnological applications.

The invention discloses a molecular marker for identifying pepper male sterility recovery traits and application of the molecular marker, and belongs to the technical field of molecular biology, the invention develops two molecular markers dCAPS and KASP for screening pepper cytoplasmic male sterility recovery traits, PR-dCAPS and PR-KASP primers developed by the invention enrich a marker library for screening the pepper male sterility recovery traits, and the molecular markers are used for identifying the pepper male sterility recovery traits. The problems that a molecular marker is narrow in application range, and more materials carrying capsicum cytoplasmic male sterility restoring genes cannot be screened out are solved. The two molecular markers dCAPS and KASP provided by the invention can be used for effectively identifying the variety, strain and single plant of pepper containing the restoring gene associated with the molecular markers, and the markers can also be used for carrying out resultant identification on the fertility restoration of pepper commercial species, so that the breeding time is greatly shortened, the working efficiency is improved, and the molecular markers have wide market application prospects.

The invention relates to a yarrowia lipolytica strain for producing tyrosol as well as a construction method and application of the yarrowia lipolytica strain, and belongs to the technical field of genetic engineering. According to the present invention, ScADH6, ScARO10F138L and D218G genes are expressed in the yarrowia lipolytica W29 [delta] KU70; the shikimic acid feedback inhibition is relieved by expressing the Y1ARO3K225L gene, the Y1ARO4K2214 gene and the Y1AR07G139S gene; pmLAAD, EcTyrAM53I and A354V genes are introduced, so that the conversion from FAD to FADH2 is promoted, and the supply of NADH (Nicotinamide Adenine Dinucleotide Hormone) in cytoplasm is enhanced; a 4-hydroxyphenylpyruvate dioxygenase (HPD) gene is knocked out; and overexpressing Y1DHS1, Y1DHS2 and Y1DHS3 genes of a shikimic acid pathway to obtain the yarrowia lipolytica strain HY5 for producing tyrosol. The invention lays a foundation for industrial production of tyrosol, and has important social significance and economic value.

The invention discloses a virus-like particle coated with an aluminum-containing metal organic framework mineralization layer and application of the virus-like particle. In order to improve the VLPs vaccine stability and immune effect, the invention synthesizes a novel aluminum-containing metal organic framework (ZAM). The ZAM can mineralize the VLPs at a high level under a mild condition to form the VLPs-ZAM nano vaccine. Taking a foot and mouth disease virus (FMDV) virus-like particle (VLPs) vaccine as an example, a heat treatment test shows that ZAM mineralization significantly improves the heat stability of FMDVVLPs, and the effect is superior to that of Al (OH) 3 and ZIF-8. The FMDV VLPs-ZAM not only has the effect of promoting APCs to take in FMDVVLPs, but also can promote antigens to escape from lysosome to cytoplasm due to the pH responsiveness of the FMDV VLPs-ZAM. A mouse immune test shows that ZAM mineralization improves the specific immune response level and stability induced by FMDV VLPs. The invention provides a new technical means for improving the stability of the VLPs vaccine and the immune effect of the VLPs vaccine.

This invention discloses a rapid method for creating wheat CMS maintainer lines based on recurrent selection and double haploid technology. The method involves crossing a CMS sterile line with a fertile cytoplasmic restorer line to obtain heterozygous fertile plants. Then, using the restorer line as the recurrent paternal parent, a backcross is performed to convert the cytoplasm to a fertile genotype. Double haploid technology is used to rapidly obtain a homozygous DH population from the backcross progeny. Finally, through testcross identification, DH plants with completely sterile cytoplasmic cytoplasm are selected, thus obtaining the target maintainer line with fertile cytoplasm and a sterile nuclear genotype. This invention further incorporates recurrent selection technology for continuous optimization of the maintainer line's agronomic traits. This invention integrates backcrossing, DH technology, and a recurrent selection system, significantly shortening the breeding cycle. The maintainer line exhibits homozygosity ≥99%, a pure cytoplasmic background, strong versatility, and applicability to wheat CMS types, providing efficient technical support for hybrid wheat breeding.

The present invention relates to modified myeloid cells or iPS or HSCs comprising a chimeric antigen receptor (CAR), or modified induced pluripotent stem cells (iPS) or hematopoietic stem cells (HSCs) comprising a CAR, the CAR comprising an extracellular antigen binding domain that binds to a tumor antigen or a tumor microenvironment (TME) antigen, a transmembrane domain, and a first intracellular signaling domain comprising the cytoplasmic tail of CD40 fused to a second intracellular signaling domain comprising the intracellular domain of CD3ζ. The present invention also relates to therapeutic uses thereof.

The invention relates to the technical field of single-cell RNA (Ribonucleic Acid) sequencing, and discloses a CellRanger single-cell RNA sequencing data analysis method, which comprises a grading treatment for cell quality screening and a tolerance mode for gene expression screening. The method provided by the invention not only strictly ensures the integrity of each functional index in the cells which are finally put in storage, but also retains part of cell information which possibly has biological significance and causes candidate marker deletion due to low overall transcriptional activity by further judging the low expression state in the to-be-determined cells. Through the method, the reliability of subsequent analysis data can be ensured, and omission of information of low-expression or rare cells due to a strict one-step standard can be reduced as much as possible.

The invention discloses a method for creating a cytoplasmic male sterile line of flowering Chinese cabbage, which comprises the following steps of: (1) selfing Zhengcheng tip leaf flowering Chinese cabbage to find a sterile plant; (2) carrying out test cross on sterile plants and sister plants appearing in the Zhengcheng sharp-leaf flowering cabbage inbred line to obtain test cross seeds; (3) planting the test-cross seeds, observing fertility and nectary conditions in a flowering period, backcrossing sterile plants and corresponding sister plants in a group with the test-cross offspring sterility rate of more than 50%, screening out a group with the sterility rate of 100% from backcross offsprings, and selfing the sterile plants in the group with the corresponding sister plants to obtain the sister plants with the infertility rate of more than 50% and the infertility rate of more than 100% in the group with anther degradation and normal pistil and nectary. Selfing the fertile sister plants to obtain offspring to form a maintainer line, and performing test crossing on the sterile plants and the maintainer line to obtain a matched sterile line. According to the method, transformation of a sterile source is not needed, time is saved, cost is low, the sterile line preserves completeness of nectary genes, nectary is normal, the podding rate is high, the maturing rate is high, and the seed yield is high.

The invention relates to the technical field of biological detection, in particular to a bHLH090 gene promoter and application in a method for detecting and counting the number of arabidopsis myrosinase heteromorph, and the sequence of the bHLH090 gene promoter is shown as SEQ ID No.1. The bHLH090 gene promoter is selected from bHLH090 genes specifically expressed in the arabidopsis myrosinase heteromorph, and the bHLH090 gene promoter can be used for detecting and counting the number of the arabidopsis myrosinase heteromorph in the arabidopsis myrosinase heteromorph. A nuclear localization fusion expression vector of a specific expression gene bHLH090 is constructed, the bHLH090 gene specifically expressed by myrosinase cells is cut in, and a marker originally expressed in cytoplasm is localized to a cell nucleus through related construction and connection operations, so that accurate marking and localization of myrosinase heteromorphic cells are realized, and the accuracy of the myrosinase heteromorphic cells is improved. Specific marking and living body observation of myrosinase heteromorphic cells are achieved, and the lethal problem of a traditional dyeing method is avoided.

The invention discloses application of cell surface markers in quality evaluation of umbilical cord mesenchymal stem cells and application of a kit for detecting the cell surface markers in quality evaluation of the umbilical cord mesenchymal stem cells. The cell surface markers are CD142, CD146, CD106 and CD362. The invention also discloses a quality evaluation method of the umbilical cord mesenchymal stem cells. The method comprises the following steps: detecting the expression condition of the cell surface markers of the cells to be detected, and evaluating the cell quality according to the expression result, the cell surface markers are CD142, CD146, CD106 and CD362, and the cell surface markers are used for detecting the cell surface markers. According to the method, accurate detection and systematic analysis of specific markers of different generations of multiple umbilical cord samples are realized, an influence rule of passage and sample individuals on marker expression is defined, and an established cell characteristic evaluation system can accurately judge the quality and functional potential of the umbilical cord mesenchymal stem cells; the scientific basis is provided for the application of the umbilical cord mesenchymal stem cells.

This invention discloses a specific molecular marker for identifying eggplant Saet CMS cytoplasmic male sterility lines and its application. The molecular marker, Saet218b, was selected by comparing the mitochondrial genomes of the eggplant Saet CMS cytoplasmic male sterility line and its maintainer line. Through bioinformatics analysis, the candidate specific gene for Saet CMS male sterility, orf218b, was obtained, and its nucleotide sequence is shown in SEQ ID NO: 1. Based on this sequence information, the molecular marker Saet218b for identifying eggplant Saet CMS cytoplasmic male sterility lines was successfully designed. The molecular marker of this invention can be used to identify eggplant Saet CMS cytoplasmic male sterility lines efficiently and with a simple procedure. The results can be used for marker-assisted breeding of eggplant Saet-derived cytoplasmic male sterility lines, improving breeding efficiency. It also provides technical support and theoretical basis for the identification of eggplant specific cytoplasmic germplasm.

The invention relates to a lung fibroblast culture method, and belongs to the technical field of cell culture. Comprising the following steps: S1, organization preparation; s2, tissue treatment; s3, primary culture: the treated tissue blocks are attached to a culture dish, a culture solution is added, the culture solution is a primary culture system, and culture is performed in an incubator for several days until the culture dish is full of fibroblasts; s4, subculture: digesting the cells subjected to primary culture by adopting conventional 0.25% trypsin, then adding a culture solution which is a subculture system for subculture, and carrying out subculture to a fourth generation; and S5, cell identification. According to the invention, primary and subculture are respectively purified and amplified by virtue of a double culture system of fibroblasts, so that the purification of target cells is facilitated, the cell quality is good, a low-serum growth additive is used as a nutrient substance, the low-serum growth additive is more easily utilized by the cells, the proliferation of the cells is facilitated, and good proliferation activity and phenotypic stability are still kept when the cells are subcultured to the fourth generation; no obvious differentiation phenomenon occurs.

The invention provides a culture medium for NK cell culture and a preparation method of NK cells. The culture medium comprises a coating solution, an activation culture medium and a proliferation culture medium, the coating solution comprises an antibody, recombinant human fibronectin and a buffer solution; the activation culture medium comprises a first cell factor, a glycogen synthase kinase inhibitor and an NK basal culture medium; the proliferation culture medium comprises a second cell factor and an NK basal culture medium. The problem that in the prior art, prepared NK cells are poor in quality can be solved, and the method is suitable for the field of cell culture.

This invention belongs to the field of plant genetic engineering, specifically disclosing a method for creating bifunctional haploid inducible lines of Brassica napus based on the BnaKNL2 gene and its application. Research revealed that the BnaKNL2 gene is a key gene affecting the fertility of Brassica napus. By knocking out the KNL2 gene in Brassica napus, bifunctional haploid inducible lines with maternal or paternal haploid induction capabilities can be created. The haploid induction rate after hybridization with different Brassica napus varieties can reach 0.45%–2.7%. Furthermore, by combining this with a cytoplasmic male sterility system and through hybridization and backcrossing, the CMS-HI line, carrying both sterile cytoplasm and haploid induction capabilities, can be rapidly created. This provides a new approach for haploid breeding of Brassica napus and the improvement and creation of CMS lines, with broad application prospects.

The invention relates to an iridium (III) complex as well as a preparation method and application thereof in head and neck cancer, and belongs to the technical field of biological medicines. The novel iridium (III) complex provided by the invention has cytotoxicity to head and neck cancer, and the cytotoxicity is better than that of cis-platinum CDDP. The iridium (III) complex acts on head and neck cancer, and can inhibit migration and proliferation of tumor cells, induce apoptosis of the tumor cells, induce CRT expression quantity increase and overturn to cell surfaces, induce HMGB1 expression quantity increase and transfer to cytoplasm and release, induce tumor cells to release more ATP, activate endoplasmic reticulum stress pathways, improve Ca < 2 + > level in the tumor cells, inhibit tumor cell apoptosis, inhibit CRT expression quantity increase and overturn to cell surfaces, induce HMGB1 expression quantity increase and transfer to cytoplasm and release and inhibit tumor cell apoptosis. The mitochondrial membrane potential of tumor cells is reduced, and the level of active oxygen is improved; tumor growth is inhibited, tumor apoptosis is promoted, tumor immunogen cell death is induced through endoplasmic reticulum stress, and tumor immune response is induced. A new choice is provided for preparing the medicine for treating the head and neck cancer.

A wheat transgenic plant carrying restorer of fertility genes specific to T. timopheevii CMS cytoplasm.

To provide a method for vector-free delivery of a gene editing composition across a plasma membrane of a cell.SOLUTION: A method for delivering a gene editing composition across a plasma membrane of a cell, comprises the steps of: providing a population of cells; and contacting the population of cells with a volume of aqueous solution, the aqueous solution comprising the gene editing composition and an alcohol at a concentration of at least about 2%, wherein the volume is a function of: (i) exposed surface area of the population of cells; or (ii) a number of cells in the population of cells.SELECTED DRAWING: Figure 1

The invention discloses a preparation method and application of cytoplast vesicles. The preparation method is realized through a dehydration method. After the cells are dehydrated and shrink, a culture medium is added for culture, a large number of vesicles with organelles such as ribosomes and mitochondria and cell matrixes can be rapidly secreted in a short time, and the vesicles can be centrifugally collected and further used for later processing. The method is convenient and rapid, is not limited by cell types, is high in yield, does not involve the use of toxic substances, and can change the functions of the cytoplast vesicles by transforming parent cells. The prepared cytoplast vesicles can be used for disease diagnosis, clinical medication and the like, have high application value and are suitable for application and popularization.

The invention relates to the cross technical field of molecular biology and parasitology, and particularly discloses a giardia living body marking method based on fluorescent protein UnaG, which comprises the following steps: firstly, optimizing and synthesizing a UnaG gene sequence according to the codon use preference of giardia; then, constructing a marker plasmid pGT-UnaG-neo on a double expression framework containing an alpha2-tubulin promoter and a gdh promoter, and introducing the plasmid into the giardia giardia trophozoite through electroporation. And carrying out resistance screening in a culture medium containing geneticin to obtain an insect strain capable of stably integrating and expressing UnaG. Through bilirubin induction, efficient maturation and green fluorescence emission of the UnaG protein in giardia cytoplasm are realized. The stably transferred strain obtained by the method has no significant difference from a wild strain in the aspects of growth curve and survival rate, and UnaG and bilirubin have no obvious toxic effect. The limitation that traditional fluorescent protein is inactivated under the anaerobic condition is broken through, and a reliable tool is provided for dynamic imaging, functional research and drug screening of giardia and other anaerobic protozoa.

A cell quality evaluation apparatus performs a process including estimation processing of estimating a quality of cells in the entirety of a cell-culture container by determining feature quantities from a plurality of images and calculating an average value of the feature quantities; derivation processing of deriving an estimation error of the quality estimated in the estimation processing, based on a variation of the feature quantities in the plurality of images and imaging information related to an area of a plurality of imaging regions; and imaging control processing of causing a imaging apparatus to perform re-imaging on at least one re-imaging region different from the plurality of imaging regions in a case where the estimation error is out of an allowable range.

The present application relates to a kind of CD69 humanization mouse model construction method, the construction method: (1) the targeting vector of expression humanization CD69 gene is constructed;(2) design and obtain sgRNA for mouse Cd69 gene;(3) the targeting vector, sgRNA and Cas9 protein are co-injected or co-electrotransferred to mouse fertilized egg cytoplasm or nucleus, and the fertilized egg is transplanted to pseudopregnant mouse, and genotype identification is carried out to pseudopregnant litter mouse, and the positive F0 mouse of correct human source fragment successfully inserted is screened;(4)F0 mouse is mated with background mouse to obtain F1 mouse, and CD69 humanization mouse model is screened out.The CD69 humanization mouse constructed in the present application has application value in immunology and other fields.

This invention relates to a method for constructing a humanized FXI mouse model, comprising: (1) constructing a targeting vector expressing the humanized FXI gene; (2) designing and obtaining sgRNA targeting the mouse FXI gene; (3) co-injecting or co-electroporating the targeting vector, sgRNA, and Cas9 protein into the cytoplasm or nucleus of mouse zygotes, and transplanting the zygotes into pseudopregnant mice, performing genotyping on the pseudopregnant pups, and screening for positive F0 mice with the correct humanized fragment inserted; (4) mating the F0 mice with background mice to obtain F1 mice, and screening for the humanized FXI mouse model. The humanized FXI mouse constructed by this invention has broad application prospects in the coagulation system, thrombotic diseases, and related drug screening.

The invention belongs to the technical field of biology, and particularly relates to an indel molecular marker linked with a radish NWB cytoplasmic male sterility (NWB-CMS) fertility restoring gene and application. Comprising first to fourth Indel marks. The size of the first Indel marker is 156 bp or 150 bp, the size of the second Indel marker is 115 bp or 109 bp, the size of the third Indel marker is 179 bp or 171 bp, and the size of the fourth Indel marker is 106 bp or 90 bp. According to the invention, a locus for controlling NWB-CMS fertility recovery of radish is excavated, and a molecular marker related to the locus is developed, so that micro-powder and sterile plants can be identified and selected more quickly and accurately, and fertile plants can be effectively eliminated, thereby accelerating the breeding process and improving the breeding efficiency.

This invention relates to the field of biomedical technology and discloses a biomarker for diagnosing polycystic ovary syndrome (PCOS) and its application. The diagnostic biomarker is circSPECC1(4), which is formed by reverse splicing and circularization of the fourth exon of the SPECCC1 gene. It has a length of 1580 nt and possesses a closed circular RNA structure. circSPECC1(4) is specifically highly expressed in ovarian granulosa cells of PCOS patients, and there is no significant difference in the mRNA expression level of its parent gene SPECCC1. circSPECC1(4) possesses RNase R nuclease resistance stability and is mainly located in the cytoplasm. Knocking down the expression of circSPECC1(4) can significantly promote apoptosis of ovarian granulosa cells and arrest the ovarian granulosa cell cycle at the G0 / G1 phase. This invention clearly demonstrates that circSPECC1(4) can serve as a specific diagnostic biomarker for PCOS, fully verifying the structural characteristics, expression specificity, and pathological regulatory function of this circular RNA, filling the gap in the existing field of PCOS diagnosis which lacks highly specific and stable molecular diagnostic targets.

The application relates to the field of stem cell biotechnology and discloses a stem cell quality evaluation system and method, a stem cell quality evaluation system, which comprises a data acquisition module, which acquires local environment data and image data of stem cells within a preset range; a morphology identification module, which obtains cell morphology data of all stem cells within the preset range through a cell morphology identification model; a graph structure data construction module, which constructs graph structure data; a scoring and labeling module, which determines the proliferation capacity, genetic stability capacity and surface marker expression capacity of stem cells after a preset time length, and obtains the quality score of the stem cells through artificial labeling; and a model training module, which trains a stem cell quality scoring model through an optimization algorithm; the cell morphology data of the stem cells are extracted through a multi-branch network of the cell morphology identification model, and the interaction between the stem cells is comprehensively considered through the stem cell quality scoring model, so that the accuracy of stem cell quality evaluation is improved.

The invention provides a real-time detection and dynamic feeding system and method for an NK cell suspension culture nutrient solution. The system comprises a multi-parameter sensing module used for monitoring the concentration of multiple components in a culture solution in real time; the intelligent decision control module is used for dynamically predicting nutritional requirements and generating a feeding strategy according to the multi-parameter data monitored in real time and based on related models and algorithms; the precise feed supplement execution module is used for injecting a nutrient solution and a pH adjusting solution as required through a multi-channel peristaltic pump according to the generated feed supplement strategy, supplementing growth factors at regular time, and is provided with a metabolic waste removal unit for removing lactic acid and ammonia metabolic waste; and the feedback optimization database is used for recording culture process data, optimizing a feeding strategy through a machine learning algorithm, and adapting to individual requirements of different cell strains. The technical problems that in the prior art, a static feeding strategy cannot respond to nutrition dynamic changes in real time, metabolic byproducts lack a real-time removal mechanism, and the cell quality is unstable due to nutrition imbalance are solved.