10 results about "Cell plasma membrane" patented technology

The invention belongs to the technical field of bioengineering, and discloses a plasma membrane localized green fluorescence overexpression vector as well as a construction method and application thereof. The cytoplasmic membrane localized green fluorescence overexpression vector contains an expression vector skeleton and an AtCBL1 gene connected to the expression vector skeleton, wherein the nucleotide sequence of the AtCBL1 gene is shown as SEQIDNO: 1, and the expression vector skeleton also carries a fluorescent protein gene. The construction method of the vector comprises the following steps: S1, carrying out enzyme digestion and recovery on an AtCBL1 gene sequence with a nucleotide sequence as shown in SEQIDNO: 1 obtained by amplification; s2, performing same enzyme digestion on the expression vector carrying the fluorescent protein gene, and recovering; and S3, connecting the AtCBL1 gene subjected to enzyme digestion with the expression vector to form a recombinant plasmid, converting a connection product into an escherichia coli competent cell, and extracting the recombinant plasmid from positive clone to obtain the plasma membrane localized green fluorescence overexpression vector.

Delivery of a payload across a cytoplasmic membrane includes providing a population of cells, contacting the population of cells with a volume of an aqueous solution. The aqueous solution includes an effective load and an alcohol at a concentration greater than 5%. The volume of the aqueous solution may be a function of the exposed surface area of the cell population, or a function of the number of cells in the cell population. Related compositions, instruments, systems, techniques, and articles are also described herein.

Crimean-Congo hemorrhagic fever virus (CCHFV) is an important human pathogen. Limited evidence suggests that antibodies can protect humans against lethal CCHFV disease, but the protective efficacy of antibodies has never been evaluated in adult animal models. Here adult mice were used to investigate the protection provided by glycoprotein-targeting neutralizing and non-neutralizing monoclonal antibodies (mAbs) against CCHFV infection. A single non-neutralizing antibody (mAb-13G8) was identified that protected adult type I interferon deficient mice >90% when treatment was initiated prior to virus exposure and >60% when administered after virus exposure. Neutralizing antibodies known to protect neonatal mice from lethal CCHFV infection, failed to confer protection regardless of IgG subclass. The target of mAb-13G8 was identified as GP38, one of multiple proteolytically-cleaved glycoproteins derived from the CCHFV glycoprotein precursor polyprotein. Robust protection required complement activity, but not Fc-receptor functionality. Consistently, it was found that GP38 previously identified as a secreted molecule also localizes to viral envelope and cellular plasma membranes. This study reveals GP38 as an important antibody target for CCHFV and lays the foundation to develop novel vaccines and immunotherapeutic against CCHFV in human.

The invention relates to the technical field of cell biology, in particular to a non-cell stimulant for amplifying NK cells derived from pluripotent stem cells. The invention provides an extraction method of an aAPC cell membrane. According to the extraction method, the amplification efficiency of aAPC on NK cells can be remarkably improved through specific cell factors or a combination of the specific cell factors; after the cells are subjected to hypotonic treatment, the cells are filtered by a centrifugal column, and the extracted cytoplasmic membrane (PM) component does not have living cells and is high in safety when being used for NK (Natural Killer) cell amplification; the PM film extraction process is simple and efficient, PM film preparation can be performed in a common laboratory with a high-speed refrigerated centrifuge, and large-scale preparation is easy; by means of the scheme (the hypotonic solution and the centrifugal tubular column), the PM extraction yield is high, 1500-2000 micrograms of PM component protein can be obtained through 5.0 E7 cells, and the whole process is short in consumed time (about 45 min can be completed).

This disclosure provides a method and apparatus for identifying enhancer activity in live cells, enabling sensitive detection of enhancer activity within live cells. The method utilizes a nano-electroporation biochip to perform highly focused perforation of the cell membrane and provides the driving force to accelerate the delivery of enhancer probes to the cell nucleus. The enhancer probes are designed using a cyclic amplification strategy, increasing the fluorescence signal by more than 100-fold within one hour. In the embodiments, the method and apparatus successfully detected the activity of CCAT1 enhancer in live cells without affecting cell viability, demonstrating the robustness and reliability of the proposed method and apparatus in detecting enhancer activity in live cells. This disclosure provides a convenient and universal platform for research on enhancer gene regulatory mechanisms in live cells and for clinical diagnostic applications.

The invention relates to the technical field of fluorescent probes, and discloses a preparation method and application of a cytoplasmic membrane covalent labeling fluorescent probe based on non-membrane permeability, and the preparation method comprises the following steps: S1, synthesizing 4-(2, 3, 3-trimethyl-3H-indole-1-onium-1-yl) butane-1-sulfonate; s2, synthesizing a 2-halogen substituted N-((1E, 2Z)-3-9 (phenylamino) allylidene) phenylamine derivative; and S3, synthesizing a target product, namely the Sulfo-Cy5-X fluorescent probe. The novel cytoplasmic membrane fluorescent probe is prepared by adopting a membrane impermeability-based cytoplasmic membrane covalent labeling principle and introducing a sulfonic acid group with high water solubility and a covalent reaction site, so that a long-time and specific labeling function on the cytoplasmic membrane is realized; according to the method, the problems of short imaging time and low water solubility of a traditional fluorescent probe are solved, and a more ideal tool is provided for in-situ observation of dynamic change of the cytoplasmic membrane.

The invention discloses a pediococcus pentosaceus D17-sourced cytoplasmic membrane vesicle and application thereof, and belongs to the technical field of microorganisms. The pediococcus pentosaceus D17 with excellent probiotic characteristics is separated from fresh excrement of Lichuan horses for the first time, the preservation number of the pediococcus pentosaceus D17 is CCTCC (China Center For Type Culture Collection) NO: M20252051, and meanwhile, the application of the pediococcus pentosaceus D17 and the cytoplasmic membrane vesicles of the pediococcus pentosaceus D17 in resisting salmonella infection is researched. Experimental verification shows that the separated and screened pediococcus pentosaceus D17 and the cytoplasmic membrane vesicles thereof have the effect of resisting salmonella infection and can promote expression of tight junction protein, and the treatment effect of the cytoplasmic membrane vesicles derived from the pediococcus pentosaceus D17 is superior to that of the pediococcus pentosaceus D17. Due to the fact that the cytoplasmic membrane vesicles are prepared through centrifugation of bacterial liquid supernatant, the cytoplasmic membrane vesicles are safer and free of drug resistance risks, and a new thought and a new approach are provided for application of the cytoplasmic membrane vesicles sourced from pediococcus pentosaceus to preparation of salmonella infection resisting products.

The application discloses a water-soluble AIE fluorescent probe, and a structure formula is shown in the following formula (I), wherein R1 and R2 are independently selected from alkyl alcohol with carbon number of 1-6; X is selected from water-soluble anions, and M is selected from water-soluble cations. The fluorescent probe has AIE characteristics, can realize rapid and efficient staining of cytoplasmic membranes, and does not need to be cleaned; the fluorescent probe has good water solubility, small cytotoxicity, and can be used in in vivo cell fluorescence imaging; the fluorescent probe has orange red fluorescence color, large Stokes shift, and strong anti-self-fluorescence interference capability.

This invention relates to a method for increasing the permeability of a cell membrane by directing at least one cell to or near a structure containing particles capable of absorbing electromagnetic radiation. These particles are present at a concentration of 0.001 vol% to 20 vol% and are embedded in the material of the structure. At least 60% of the particles present in the structure are embedded in the material such that the shortest distance L between these particles and the free surface S of the structure is 1 nm to 500 nm. The invention also relates to a structure suitable for permeating cells during photothermal processes and the use of said structure.

The application discloses a water-soluble cationic fluorescent probe dye and application thereof in cell membrane staining. The fluorescent probe has good water solubility, can be used in in-vivo cell fluorescence imaging, realizes rapid and efficient staining of cell plasma membranes, and does not need to be cleaned. The fluorescent probe has an orange red fluorescence color, has a large Stokes shift, and has strong anti-self-fluorescence interference capability.