45 results about "Bioluminescence" patented technology

The invention discloses polyketide synthase or a polyketide synthase mutant and application thereof, and belongs to the technical field of gene engineering. The polyketide synthase is AspiPKS4, AspiPKS5, AspiPKS6 and AspiPKS7 from cyathea spinulosa, FhPKS2, FhPKS3, FhPKS4, FhPKS5 and FhPKS6 from ficus himalayana, and CrPKS from floating fern. The mutant is obtained by carrying out single point mutation on the second amino acid of AspiPKS7 or deleting an N-terminal redundant sequence. The polyketide synthase or the mutant is used for replacing NnHisspS in an original FBP luminescence system, and the biological self-luminescence level can be remarkably improved. The invention further provides a simplified and improved FBP system which comprises the polyketide synthase or the polyketide synthase mutant, CPH, H3H and Luz, and the application convenience and the application luminescence capacity of the FBP system are improved.

The invention discloses NnLuz and NnH3H protein mutants and an application of the NnLuz and NnH3H protein mutants in enhancing the bioluminescence intensity of fungi or plants. According to the method disclosed by the invention, core enzymes NnLuz and NnH3H in a fungus bioluminescence pathway are optimized through rational design, and the catalytic efficiency and the luminous intensity of a bioluminescence system are remarkably improved. Experiments prove that 1) the luminous intensity of the NnLuz [delta] N19v2 (I103T / L167K double mutant) in yeast and plants is remarkably superior to that of a reported NnLuzv4 mutant and is 3-4 times that of the reported NnLuzv4 mutant, 2) the catalytic efficiency of the NnH3Hv3 (K307T mutant) is improved by more than 2 times, the luminous stability is remarkably enhanced, and 3) when the NnLuz [delta] N19v2 and the NnH3Hv3 are cooperatively expressed, the luminous intensity of the system reaches the optimal level, and the self-luminous plant with remarkably enhanced luminescence is created.

The present invention relates to a method for screening compounds for bactericidal activity using thermostable luciferase and based on a real-time bioluminescence measurement. The present invention further relates to a method for determining the sensitivity of a bacterial sample originating from a subject suffering from a bacterial infection to a group of known antibiotics and to a method for assessing the minimum inhibitory concentration (MIC) of a bactericidal compound.

The invention belongs to the technical field of microbial nucleic acid detection and bioluminescence analysis, and discloses a bioluminescence nucleic acid detection method and kit based on DNA-RNA hybrid specific recognition. The kit comprises a streptavidin coated solid phase carrier, a biotinylated DNA capture probe, a signal enhancement DNA, a luminescence report fusion protein and a luminescence substrate thereof, the luminescent reporter fusion protein comprises a DNA-RNA hybrid binding domain and a luminescent enzyme domain. According to the method, target RNA is captured through a solid phase, a DNA-RNA hybrid is formed, the luminous report fusion protein is specifically combined with the DNA-RNA hybrid, a bioluminescence signal is generated after a luminous substrate is added, and the target RNA is detected. In the embodiment, an amplification step is not adopted, so that the method has the advantages of simplicity and convenience in operation, rapidness in detection, lower background and better specificity, and can be used for rapid detection of microorganisms.

The invention relates to the technical field of food hazard factor detection, in particular to a food hazard factor detection method and system based on biomimetic hydrolase catalytic fluorescence generation. Comprising the following steps: S1, obtaining a turbidity characteristic value, background fluorescence intensity and a fluorescence quenching coefficient of a to-be-detected sample solution; s2, based on the turbidity characteristic value and the fluorescence quenching coefficient, calculating to obtain a robustness index; s3, according to a comparison result of the robustness index and a preset qualified threshold value, through a preset closed-loop feedback control logic, determining a working gain used for total fluorescence intensity signal acquisition; s4, performing dynamic signal correction processing including gain normalization to obtain a normalized apparent fluorescence signal; s5, based on the normalized apparent fluorescence signal, calculating to obtain an apparent catalytic rate; and S6, determining the concentration of the target hazard factor according to the apparent catalytic rate, the turbidity characteristic value and the fluorescence quenching coefficient. According to the method, the accuracy and the reliability of detecting the target hazard factors in the complex matrix are remarkably improved.

The invention provides application of a phellinus polyketide synthase gene SvPKS1 in promoting bioluminescence of fungi or plants, the coding sequence of the SvPKS1 is shown as SEQ ID NO.1, and the fungi are fungi for producing caffeic acid. According to the application, the SvPKS1 and other genes (nnH3H, nnLuz and nnCPH) of an FBP fungus luminescence pathway are co-expressed in fungi or plants by utilizing a transgenic technology, so that the fungi or plants are promoted to emit light biologically. The invention discloses co-expression of the phellinus polyketide synthase gene SvPKS1 and fungus luminescence pathway genes nnH3H, nnLuz and nnCPH for the first time, promotes fungus or plant bioluminescence, and provides a key candidate enzyme gene for promoting fungus or plant sustainable luminescence.

1. The name of the design product: adenosine triphosphate biological fluorescence detector. 2. The use of the design product: adenosine triphosphate biological fluorescence detector is used to quickly detect the amount of microorganisms and other biological residues in the sample, and to judge the health condition. It is suitable for monitoring the key control points in food and beverage production process, and for real-time sampling monitoring by medical system and health supervision institutions. 3. The design points of the design product: in shape. 4. The picture or photo that best indicates the design points: perspective drawing.

The invention provides an adjustable bioluminescence imaging lighting device based on a fiber Bragg grating. The adjustable bioluminescence imaging lighting device comprises a light source; the first lens unit is used for converting light rays emitted by the light source into parallel light rays; the rotary light splitting unit is used for enabling all the parallel light rays to penetrate or splitting the parallel light rays into transmission light and reflected light; the second lens unit is used for transmitting the focused light and inputting the focused light to the first fiber bragg grating through the first coupler; the first fiber bragg grating is used for reflecting the narrow-band light of the first preset wave band and outputting the narrow-band light through the output end of the first coupler; the third lens unit is used for focusing the reflected light and inputting the focused reflected light to a second fiber bragg grating through a second coupler; the second fiber bragg grating is used for reflecting narrow-band light of a second preset wave band and outputting the narrow-band light through the output end of the second coupler; and the third coupler is used for mixing the input light rays of the first / second preset wave bands and then outputting the mixed light rays to the light source input end of the endoscope. The device can be used for clinical diagnosis and detection.

The application discloses a method for identifying Klebsiella pneumoniae gene circadian rhythm expression by using a bioluminescence reporting system. The application constructs a recombinant bioluminescence Klebsiella pneumoniae driven by a metabolic gene malate:quinone oxidoreductase mqo promoter (Pmqo), and directly observes the change rule of bacterial gene expression under entrainment and free-running conditions of temperature cycle by monitoring the luminescence characteristics. The luxCDABE gene operon comprises genes for coding luciferase and substrate synthesis enzyme, wherein luxA and luxB code for heterodimer luciferase, luxC, luxD and luxE synthesize aldehyde as a substrate for the luciferase luminescence reaction. The luciferase catalyzes the oxidation of aldehyde to generate visible blue-green light with a wavelength of 450-490 nm. The Lux bioluminescence reporting system has the advantages of not needing to add an exogenous substrate and not affecting the normal physiological functions of the host, and can be applied to the identification of Klebsiella pneumoniae gene circadian rhythm expression.

This specification provides systems, methods, and compositions for bioluminescence-induced photocatalytic activation of molecular entities in a proximity-dependent manner, which can be operated within biological systems. In particular, this specification provides activatable molecular entities, systems thereof, including bioluminescent proteins or complexes, their luminescent substrates, photocatalysts, and activatable molecular entities, which incorporate photoresponsive moieties that limit their activity, as well as methods for catalytically activating activatable molecular entities via bioluminescence-induced catalysis.

The invention provides a synchronous quantitative detection method of cholate-resistant gram-negative bacteria in different active states, which comprises the following steps: (1) adding a capture probe into a bacteria solution to be detected, carrying out illumination culture to obtain a compound solution, and centrifuging the compound solution to obtain a supernatant and a precipitate resuspension; (2) adding a signal probe into the resuspension, and carrying out illumination inactivation and incubation to obtain a sandwich compound solution; (3) calculating to obtain the total bacterial count and the number of bile salt-resistant gram-negative bacteria in the living body; (4) calculating to obtain the quantity of bile salt-resistant gram-negative dead bacteria; and (5) calculating to obtain the quantity of the cholate-resistant gram-negative sublethal bacteria. The synchronous quantitative detection of the bile salt-resistant gram-negative bacteria in different active states adopts a Raman and bioluminescence dual-signal detection method, and can realize rapid and sensitive detection of living, dead and sub-lethal bile salt-resistant gram-negative bacteria.

The invention belongs to the technical field of biological living body imaging, and provides a multi-modal fusion three-dimensional imaging device, which comprises a first box body provided with a main camera shooting unit, an auxiliary camera shooting unit, a 3D camera shooting unit and a first light source; the second box body is arranged in the first box body; a movable first top plate is arranged on the base; the objective table is arranged in the second box body, the first top plate is located between the main camera shooting unit and the objective table, the objective table and the 3D camera shooting unit can rotate relatively, the 3D camera shooting unit and the auxiliary camera shooting unit are obliquely arranged relative to the objective table, and the first light source is used for providing a needed light source for bioluminescence imaging; the main camera shooting unit and the auxiliary camera shooting unit are used for shooting multi-angle two-dimensional bioluminescence images, the 3D camera shooting unit is used for scanning and shooting three-dimensional model contour data, the X-ray mechanism is used for shooting two-dimensional X-ray skeleton images of a measured living body, and registration is carried out through images of multiple modes, so that an accurate three-dimensional model of the measured living body is obtained.

The invention discloses an application of a NanoLuc luciferase double-subunit complementary system in construction of an experimental animal model. The genome of the experimental animal model contains a NanoLuc enzyme large subunit LgBiT encoding gene and stably expresses LgBiT; correspondingly, a tested object used for invading the experimental animal model or connected into the experimental animal model is in fusion connection with NanoLuc luciferase small subunit HiBiT polypeptide serving as a label or a NanoLuc luciferase small subunit HiBiT coding gene serving as a reporter gene, and LgBiT and HiBiT are specifically combined in the experimental animal model and recombined to form the NanoLuc enzyme with catalytic activity. A strong bioluminescence signal is generated under the action of a furazine substrate, so that high-sensitivity, non-invasive and real-time dynamic imaging of in-vivo distribution, loading capacity and diffusion path of a detected object is realized. The invention also discloses a gene editing system for constructing the LgBiT transgenic mouse.

Provided herein are systems and methods for the detection and quantification of a target protein via bioluminescence and / or bioluminescent resonance energy transfer (BRET). In particular, provided herein are assay methods using a multi-component biochemical system that assembles into a detectable in vitro chemical complex at a protein target of interest.

This invention relates to a recombinant construct comprising: one or more TMV Ω enhancers; and a group of fluorescent factors configured to enable an organism or its tissues or cells to emit bioluminescence; the recombinant construct is configured to increase the biosynthetic content of caffeic acid in an organism or its tissues or cells, or to increase the bioluminescence intensity of an organism or its tissues or cells. This application, by introducing TMV Ω enhancers, achieves precise regulation of these two key genes, significantly improving bioluminescence intensity and solving the problems of poor targeting and limited synergistic effects of enhancer applications in existing technologies.

We will realize a bioluminescent lighting system that uses bioluminescence as the light source. [Solution] A bioluminescent lighting system 100 comprises a tank 10 for storing a liquid containing microorganisms with bioluminescent capabilities, a light-transmitting pipeline 20 extending to a predetermined facility as a flow path for the liquid containing microorganisms and connected at both ends to the tank 10, a pump 30 for circulating the liquid containing microorganisms stored in the tank 10 through the pipeline 20, and a control device 40 for adjusting the liquid containing microorganisms flowing through the pipeline 20. The control device 40 controls the operation of the pump 30 to adjust the flow rate of the liquid containing microorganisms and performs a process to switch between a lighting mode that utilizes the bioluminescence of microorganisms released as the liquid containing microorganisms flows through the pipeline 20 as illumination light, and a culture mode in which microorganisms are cultured in at least the tank 10.

The invention relates to the field of whole-cell biosensors, and provides a whole-cell biosensor array and an early warning model for monitoring the mildewing degree and aflatoxin content of corn peanuts. The Escherichia coli cell comprises a promoter related to the response of a volatile organic compound released by Aspergillus flavus pollution, the promoter is operably connected with a reporter gene, and an expression product of the reporter gene is used for generating a bioluminescence signal which can be detected. Through the whole-cell biosensor and the array thereof, related volatile organic compounds after corn peanuts are infected by aspergillus flavus can be accurately responded, so that rapid and high-sensitivity monitoring on aspergillus flavus mildew and toxin pollution is realized. Besides, the whole-cell biosensor array is combined with a machine learning regression model, so that the aspergillus flavus infection degree can be quantified, and the aflatoxin content can be predicted.

Provided herein are systems and methods for the detection and quantification of a target protein via bioluminescence and / or bioluminescent resonance energy transfer (BRET). In particular, provided herein are assay methods using a multi-component biochemical system that assembles into a detectable in vitro chemical complex at a protein target of interest.

Disclosed are ATP-independent bioluminescent reporter systems for enhanced in vivo imaging. Also disclosed is a modified luciferin, potassiorin, comprising a potassium-binding moiety. Also disclosed is an engineered luciferase, BRIPO, optimized for synergistic interaction with potassiorin. A further version of the disclosed system produces bioluminescence signals responsive to physiological potassium concentrations. In further instances, the system enables real-time monitoring of K+ dynamics in live cells, tissues, and animals. A further embodiment includes applications in imaging neuronal activity, studying ion flux, and developing bioluminescent indicators for diverse analytes. The disclosed system addresses limitations of traditional imaging methods, offering improved sensitivity and biocompatibility.

The invention relates to the technical field of biology, in particular to a construction method and application of a helicobacter pylori luciferase bacterial strain, the helicobacter pylori luciferase bacterial strain is obtained by introducing a Nanoluc gene into a helicobacter pylori genome, and the bacterial strain can efficiently express luciferase. The fluorescent probe can interact with a synthetic substrate furimazine to initiate lasting and bright bioluminescence, can be directly used for living body imaging, and is used for animal model construction and vaccine and therapeutic drug evaluation in the helicobacter pylori vaccine research and development process. Animal experiments prove that the helicobacter pylori luciferase strain interacts with furazine in the stomach of a living animal to emit bright and lasting bioluminescence, and is used for observing sustainable infection of the living animal. The method lays a foundation for further evaluation of helicobacter pylori vaccine protectiveness, and plays an important role in development and application of prevention and treatment vaccines and drugs.

The invention relates to the technical field of biochemical detection, and discloses a surface cleanliness detection premixing reagent based on a bioluminescence method and a preparation method thereof. Each 100 mL of the ATP detection premixing reagent comprises the following components: 0.25 to 3 mg of a substrate, 3 to 10 mL of luciferase, 0.06 to 0.12 g of a magnesium salt, 15.932 to 36.155 g of a protection component, and 0 to 100 [mu] g of a fluorescence enhancer. According to the ATP detection premixing reagent provided by the invention, the technical bottleneck that luciferase and a fluorescein substrate are easy to inactivate in a mixed state is overcome by introducing a protective component consisting of the non-reducing disaccharide, the bovine serum albumin, the antioxidant and the metal ion chelating agent, and the reagent has the advantages of high sensitivity and high sensitivity after being stored for 30 days at 4 DEG C; the relative activity of the ATP detection reagent can still be kept at 90% or above of the initial activity, the performance attenuation is far lower than that of an existing product, and the dependence of the ATP detection reagent based on a bioluminescence method on subzero 20 DEG C deep freezing preservation is eliminated.

The invention discloses a method for improving the self-luminous intensity of a plant based on energy optimization, which is characterized in that a lipoic acid protein ligase A gene LplA is expressed in a self-luminous plant containing a fungus bioluminescence system FBP, so that a transgenic plant of which the luminous intensity is remarkably improved is obtained. The LplA gene is introduced to optimize and modify an energy metabolism network and enhance the efficiency of energy regeneration in organisms, the strategy basically has no influence on the substrate content of a fungus bioluminescence system, and bioluminescence enhancement is finally realized by increasing the energy and cofactor content required in a fungus bioluminescence pathway. The energy bottleneck in a fungus bioluminescence system is broken through, and an innovative solution is provided for creating efficient self-luminous plants.

This invention belongs to the field of immunoassay detection and discloses a ZrMPNs-NlucmAb bioluminescent signal probe and its application. The probe consists of a ZrMPNs signal carrier and its electrostatically adsorbed bioluminescent enzyme-labeled monoclonal antibody NlucmAb. This invention utilizes the ZrMPNs-NlucmAb probe to construct a bioluminescent test strip (BLFA), which requires no external light source and achieves a high luminescent signal in a furazine (FZ)-Nluc system, thus enabling sensitive detection of OTA. The limit of detection (LOD) of BLFA is 0.02 ng / mL, which is 9 times more sensitive than traditional gold nanoparticle-based LFAs. The cross-reactivity rates of this BLFA to OTB and OTC are 122.2% and 111.1%, respectively, and it does not recognize other common fungal toxins, exhibiting good specificity. Its application in the detection of OTA in cereals shows great promise.

Described herein are fusion proteins and methods of using those fusion proteins for the detection and identification of analytes (e.g., multimeric proteins expressed by pathogenic organisms or mammalian cells). In some embodiments, it includes a plurality of pairs of fusion proteins distinct for one analyte wherein each pair comprises i) a first fusion protein comprising one or more binding domains fused to a first portion of a reporter protein, and ii) a second fusion protein comprising one or more binding domains fused to a second portion of the same reporter protein, wherein the first and second portions of the reporter protein, when together, comprise the reporter protein in its entirety and restore its function, wherein the first fusion protein and / or the second fusion protein further comprises a fluorophore, and wherein the fluorophore for each pair has a distinguishable emission spectrum.

The invention discloses a rapid and accurate detection method for pathogenic bacteria. The method comprises the following steps: immobilizing bacteriophages on the surfaces of magnetic nanoparticles through physical adsorption or chemical reaction, selectively enriching pathogenic bacteria in a complex sample by adopting the bacteriophage functionalized magnetic nanoparticles, quickly cracking the pathogenic bacteria on the surfaces of the magnetic nanoparticles by using CTAB (cetyltrimethyl ammonium bromide), releasing ATP in the bacteria, and quantitatively analyzing ATP by using bioluminescence. Further calculating the content of the pathogenic bacteria; the detection method is free of enrichment and amplification, can complete capture, enrichment and detection of pathogenic bacteria within 20 minutes, and can specifically detect viable bacteria of target bacteria in various complex matrix (such as food) samples due to the fact that bacteriophages are adopted as recognition molecules.

Provided herein are isolated polynucleotide encoding modified click beetle luciferase polypeptides that have enhanced luminescence and longer wavelength near-infrared signals. The disclosure also relates to near-infrared bioluminescence systems that include said modified click beetle luciferase polypeptides and novel luciferin derivatives, as well as methods of using said modified click beetle luciferase polypeptides and bioluminescence systems.

A bioluminescent single photon bioreactor for performing absolute quantification of light-producing activity by enzymes includes: a bioreactor that produces a bio-electronic signal; an electronic sensor that receives the bio-electronic signal and produces an electrical transduction signal; and an analyzer that receives the electrical transduction signal and absolutely quantifies light-producing activity by enzymes from the electrical transduction signal, such that the absolute quantification is accomplished quantum mechanically by determination of a second order autocorrelation function.