135results about How to "Simple screening method" patented technology

The invention discloses a recombinant vector and a non-GMO gene editing plant screening method. An initial carrier in the recombinant vector is a CRISPR/Cas9 carrier which contains sgRNA gene, Cas9 gene and screening label gene and is used for plant gene editing, the recombinant vector carries a BelRNAi expression element, and a hairpin RNA interference fragment interfering with bentazone resistance gene is generated through transcription of the BelRNAi expression element. By introducing the BelRNAi expression element to the initial carrier and conducting bentazone herbicide screening on future generation plants, offspring with target gene mutated are reserved, and it is also ensured that the mutated offspring do not contain transgenic fragments; screening method is cheaper, simpler and more effective.

The invention discloses a distributed optical fiber sensing signal mode identifying method and system. The distributed optical fiber sensing signal mode identifying method includes steps of acquiring samples, and acquiring several groups of samples for excitation signals of each behavior, so as to set up a sample library; roughly screening samples, screening the acquired sample library to remove samples high in discreteness; finely screening the samples, training to generate a classifier used for signal mode identification by means of the screened sample library and storing; identifying signal modes, calculating characteristic vectors for a newly triggered and read section of signal data and subjecting the characteristic vectors to calculation in the stored classifier to obtain mode identification results. The distributed optical fiber sensing signal mode identifying method and system can effectively identify invasion or damage signals in case that signals are carried with noise or distortion, and in the meantime, mistaken reports are reduced.

The invention belongs to the technical field of biological pesticides, discloses a Streptomyces l avendulae xjy strain which has been collected in the general microorganism center of CCCCM in 8.16.2007 with the collection number: CCMCC No.2130. The invention also discloses a separation and authentication method for the strain, a preparation method for antibiotic active substances of the Streptomyces l avendulae xjy strain, authentication for analysis structures of the active substances, prevention and cure experiments for metabolic active substances of the xjy strain against leaf muld of tomato, powdery mildew of cucumber and phytophthora blight of pepper, and growth-promoting experiments for the growth of the strain. The antibiotic active substances secreted by Streptomyces xjy strain is mainly nucleosides antibiotic having the characteristics of persistent effect and no medicinal hazard as well as having the effects of obviously preventing and curing vegetable diseases and promoting the growth to a certain extent.

The invention discloses a site selection method of a wireless base station, and aims at providing the site selection method of the wireless base station which realizes a maximum coverage area by using minimum available base station candidate points. The site selection method of the wireless base station comprises the steps of firstly numbering all the available base station candidate points in a two-dimensional coordinate graph based on parameter information of the available base station candidate points to obtain a two-dimensional coordinate graph; then if actual coverage area of all the base station candidate points in a target region is equal to the theoretical maximum cover area of all the base station candidate points, regarding all the base station candidate points as base station setting points; then if removal of one selected base station candidate point has not influence to the actual coverage area of the target region, cancelling the base station candidate point; and finally, if the base station distance of two base station candidate points is smaller than 1.5 times of the coverage radius of the base station candidate point, cancelling one of the two base station candidate points, which has bigger overlapping degree with the coverage regions of other base station candidate points.

The invention discloses acinetobacter baumannii, and a screening method and an application thereof in degradation of an azo dye Congo red. The stain is named as acinetobacter baumannii YNWH226, is preserved in China general microbiological culture collection center, and has the preservation number of CGMCC No.7922. The invention provides the new acinetobacter baumannii and the application thereof in degradation of the dye. Especially, the acinetobacter baumannii can effectively degrade the azo dye Congo red, has quite good decoloration and mineralization effects on Congo red wastewater, can completely mineralize the Congo red under a simple aerobic condition, and has great significance on governance of dye pollution and study on aerobic degradation of the azo dye.

The invention belongs to the field of traditional Chinese medicines. The screening method of an alpha-glucosidase inhibitor comprises the following steps of: (1) establishing in vitro screening models; establishing in vitro screening models of alpha-glucosidase and a substrate PNPG (Para Nitrobenzene-Beta-D-Galactose Pyranoside); (2) establishing a component library: selecting traditional Chinesemedicines, systematically extracting each traditional Chinese medicine with various (more than two kinds) solvents according to a similar compatible principle according to the polarities from low to high, wherein multiple extracting parts can be obtained for each traditional Chinese medicine by adopting the method; and (3) screening: blending all the extracting parts of the traditional Chinese medicines into a solution with a certain concentration, adding the solution to a reaction model, and calculating the inhibition rate. The screening method is simple, reasonable and easy to operate and can be used for finally screening out traditional Chinese medicines and traditional Chinese medicine components having obvious inhibition effects on the alpha-glucosidase. In addition, the screened traditional Chinese medicines and the traditional Chinese medicine components can be used for developing products aiming to inhibit the alpha-glucosidase and particularly used for developing drugs for reducing the blood sugar and has industrialization significance.

The present invention belongs to the field of biological pesticide technology, and includes four aspects: 1. separation and identification of Acinetobacter baumannii strain LCH0606; 2. preparation and application in preventing and controlling plant diseases of Acinetobacter baumannii liquid; 3. preparation and disease preventing application of Bauman bacterin; and 4. effective component analysis of Bauman bacterin. Acinetobacter baumannii liquid and Bauman bacterin have main effective component of Iturin A, and possess broad bactericidal spectrum, less germ resistance, no harm to plant, safety to animal, environment friendship and other advantages. In addition, Acinetobacter baumannii liquid and Bauman bacterin have certain germination promoting effect on rice seed.

The invention provides an enzyme-producing microorganism for hydrolyzing D,L-pantoyl lactone and application and a sifting method of the enzyme-producing microorganism. The method specifically comprises the steps of using an enzyme-producing microorganism strain to hydrolyze D,L-pantoyl lactone, then acidizing D,L-pantoyl lactone to form a converting solution, and conducting high-performance-liquid chromatography chirality online detection on the formed converting solution to quickly sift out strains which have the catalytic activity and stereoscopic selectivity on hydrolyzing D,L-pantoyl lactone. Through the directed sifting method, it is found that a bacterial strain Fusarium verticillioides CGMCC NO.14552 has an obvious and unique advantage in the aspect of hydrolyzing D,L-pantoyl lactone, the hydrolyzing conversion rate of 1-70% D,L-pantoyl lactone within 5-10 h can reach 35% or above, and the ee value of a target product D-pantoyl lactone is larger than 98%.

The invention provides a strain of stenotrophomonas sp., which is named as stenotrophomonas F6, and has a preservation number of CGMCC No.6547. Two active algicidal substances, namely cyclo(glycine-proline) and hydroquinone, are separated from the metabolism products of the stenotrophomonas F6, and then the substances are purified and identified. The half effective concentration of the cyclo(glycine-proline) on inhibiting microcystis aeruginosa 9110 is 9.5 mg/L, and the cyclo(glycine-proline) has no inhibiting effect on synechococcus BN60. The half effective concentrations of hydroquinone on inhibiting microcystis aeruginosa 9110 and synechococcus BN60 are 0.96 mg/L and 5.6 mg/L. The stenotrophomonas F6 and the active algicidal substances (cyclo(glycine-proline) and hydroquinone) can be applied to the development and production of novel algicide, and finally are applied to the control of cyanobacterial bloom in fresh water.

The invention discloses a Paecilomyces thermophila mutant strain and a mutation induction method and application thereof. The mutation induction method includes: Paecilomyces thermophila screened from high-temperature-fermented compost and used for producing high-temperature-resistant and acid-resistant glucose oxidase is used as the original strain, protoplast ultraviolet and ethyl methane sulfonate compound mutation induction is performed, and the screened strain is subjected to budding spore diethyl sulfate-microwave compound mutation induction to obtain the Paecilomyces thermophila mutant strain. The Paecilomyces thermophila mutant strain is preserved in China General Microbiological Culture Collection Center on January 11th, 2016, and the preservation number of the mutant strain is CGMCC No. 13182. Compared with a conventional mutation induction method, the mutation induction method has the advantages that the method is high in efficiency and simple in fast in screening, the productivity of the mutant strain is high, the yield of the mutant stain is 277.65% of that of the Paecilomyces thermophila original strain, fermentation liquor enzyme activity reaches 185U/ml, production cost is lowered, and the glucose oxidase produced by the mutant strain is good in high-temperature resistance and acid resistance and can be used as feed enzyme applied to feed industry.

The invention relates to a relay individual working life predicting and screening method based on early life performance. The relay individual working life predicting and screening method based on early life performance is characterized by comprising the following steps that under an environment temperature C1, the performance parameters C ={C2,C3, ...,Cn} and the working life D of a relay sample at the early life are acquired according to the service life test of the relay sample; a relay individual working service decision table is established based on a rough set theory and an equal probability criterion by using the C as condition attributes and using the D as decision attributes; a set of association rules with respect to the early life performance and the service life of relay individuals are extracted by the aid of attribute reduction; afterwards, for the individuals of the same specifications, the service life of the individuals can be predicted by only measuring the performance parameters of the individuals at the early life, and therefore relays can be screened. According to the method, when the working life of the relay individuals are predicted and products are screened, only the early life performance of the relay individuals is needed, no more information is needed, the time and cost for acquiring information are greatly saved, the calculation is small, and the effective service life of the relays can also not excessively shortened.

The invention discloses application of a pig SNP (Single Nucleotide Polymorphism) molecular marker in screening of reproductive traits and breeding of pigs, wherein the reproductive traits are total litter size traits and live litter size traits; the nucleotide sequence of the molecular marker is as shown in the sequence table SEQ ID NO.1; and T/C base mutation exists at the 70bp position of the sequence. The invention further provides a primer pair used for amplifying the molecular marker and a method for breeding by utilizing the molecular marker. The invention discovers that one SNP molecular marker in the pig OLR1 gene is related to the total litter size traits and the live litter size traits of pigs for the first time; therefore, the SNP molecular marker can be used for marker assisted selection breeding of pigs; namely, the invention provides new use of one SNP molecular marker in the pig OLR1 gene in marker assisted breeding of the reproductive traits of pigs; early screening ofthe reproductive traits of pigs is realized; and a screening method is simple and rapid.

The invention belongs to the field of biodegrading pesticide residue, and in particular relates to a method for screening degradation bacteria strains by taking high-efficiency cyhalothrin as a substrate. The screening method is a method for obtaining degradation bacteria which grow by taking the high-efficiency cyhalothrin as a unique carbon source through enrichment and domestication, screening, separation and purification, and reinforcement of acquired bacteria samples. The method can high efficiently and conveniently screen target active degradation bacteria, and provides strain resource for preparing related degradation bacteria agent and enzyme preparation.

The invention discloses a cordyceps militaris fruiting body development stage internal reference gene, a primer, a screening method and application, wherein the internal reference gene is TUB, and thenucleotide sequence is SEQ ID NO: 1. The nucleotide sequences of the forward and reverse primers of TUB are SEQ ID NO: 13-14. The screening method comprises the following steps: (1) selecting twelveinternal reference genes as candidate internal reference genes, and designing primers; (2) inducing cordyceps militaris fruiting body development, extracting total RNAs at different development stages, measuring RNA concentration, carrying out reverse transcription on the RNA to synthesize cDNA, taking the cDNA as a template, and analyzing the candidate internal reference genes by using real-timefluorescence quantitative PCR; and (3) carrying out Ct value analysis on the real-time fluorescence quantitative PCR data to screen out stable internal reference genes. According to the application, the TUB is used as the internal reference gene to detect the relative expression levels of target genes at different development stages. The stability of the internal reference gene is good; and the method is simple, rapid and low in cost.

The invention provides a random peptide library, wherein any one fusion protein in the random peptide library comprises a label for purifying the fusion protein, a labeling protein and random polypeptide, wherein the labeling protein is linked to the end of the carboxyl of the label, the random polypeptide is linked to the end of the carboxyl of the labeling protein. The invention also provides a method for constructing the random polypeptide library and a method for screening cells from the library to penetrate polypeptides.

The invention provides a rapid high-flux screening technology of beta-glucosidase producing strain based on a 96-orifice plate, and especially a method for adopting pNPG in high-flux screening of beta-glucosidase. The method comprises specific steps of: (1) strain reactivation; (2) reacted bacterial liquid amplification; (3) color reaction; (4) absorbance determination, and the like. According to the invention, beta-glucosidase producing microbe strains can be mined by using a pure culture isolation technology and a gene library establishing technology. Compared to a currently reported esculin plate method and a fluorometric method used in beta-glucosidase screening, the method for adopting pNPG in beta-glucosidase screening provided by the invention has advantages of convenient operation, high speed, accurate result, low price, and the like.

The invention relates to the technical field of biological medicine, and in particular, relates to an application of an anti-NMDAR autoantibody as a congenital megacolon diagnostic marker. It is found that the anti-NMDAR autoantibody has the advantages of being easy to operate, free of intervention, high in flux and low in cost in children congenital disease diagnosis, and the defect that plasma diagnosis is not available in the prior art is overcome. The method has good diagnosis sensitivity and specificity, the AUC value is 0.9046, the optimal limit corresponding sensitivity is 93.75%, and the specificity is 81.58%. Besides, children suffering from the congenital megacolon are represented as vomiting, abdominal distension, constipation, enteritis and the like, the children suffering from the symptoms are large, the children suffering from the congenital megacolon account for a small amount, and a simple and effective screening method is needed. When the method is used for disease screening, the sensitivity is 100%, the specificity is 78.95, and the method can be used for disease screening.

The invention discloses a solid-phase plate screening method of heat-resistant dextranase recombinant strain, which comprises the steps as follows: a nylon membrane is sterilized at the temperature of 121 DEG C for 15 minutes; E.coli JM109BL21(DE3)pET- 20b-Cel12B is cultured in 5ml of LB liquid culture medium for 6 hours; the LB solid culture medium is coated with 4mul of IPTG with the concentration being 5mug/mL and keeps upright at the temperature of 37 DEG C for 2-3 hours; an LB solid plate is coated with strain solution, and cultured at the temperature of 37 DEG C for 15 hours; the plate is kept at the temperature of 4 DEG C for 1-2 hours; the nylon membrane is placed on the plate to photocopy colony; the colony surface affected by the nylon membrane is upwards placed on the LB solid plate and inversely placed and cultured at the temperature of 37 DEG C for 4-5 hours; the nylon membrane is soaked in the mixture of cell wall cracking solution, cell membrane cracking solution and equilibration buffer solution for 30min; the colony surface of the nylon membrane is downwards laid on a screening plate, and kept at the temperature of 70 DEG C for 3 hours; the nylon membrane is uncovered and the screening plate is developed by 0.1% of Congo red for 15min, and decolored by 1mol/L of NaCl for 15min to select transparent circle strains. The method has the advantages of being simple, saving time and avoiding trouble, high screening efficiency and good screening effect.

The invention discloses a method for obtaining an enzyme mutant with high expression, a high activity and high stability, and belongs to the technical field of genetic engineering. By means of a functional polypeptide library which is constructed on the basis of SAPs, the whole process of obtaining the enzyme mutant is quick, efficient and convenient; meanwhile, compared with wild-type alkaline pectinase, the activity of extracellular amylase of the fused enzyme mutant of alkaline pectinase is maximally improved by 15.32 times, the stability is maximally improved by 3.86 times, and the specific enzyme activity is improved by 2.55 times; the activity of intracellular amylase of the fused enzyme mutant of lipoxygenase is maximally improved by 2.49 times, the stability is maximally improved by 3.13 times, and the specific enzyme activity is improved by 0.9 time; the activity of extracellular amylase of the fused enzyme mutant of asparaginase is maximally improved by 2.25 times, the stability is maximally improved by 3.56 times, and the specific enzyme activity is maximally improved by 1.34 times.

The invention provides an ADP-glucose pyrophosphorylase mutant and a screening method and application thereof. The screening method includes the steps of firstly, synthesizing first chain cDNA; secondly, performing PCR amplification; thirdly, building AGPase large and small subunit cDNA prokaryotic expression vectors; fourthly, performing active screening. The method has the advantages that distant hybridization is performed on AGPase large subunit genes and small subunit genes from different species, and the recombinant gene expression vectors are obtained in a highly-controllable manner; escherichia coli glgC mutant strains are converted and inoculated to a Conberg enrichment solid culture medium, and iodine-potassium iodide dyeing is used to screen the high-activity ADP-glucose pyrophosphorylase mutant. The screening method is simple and practical and stable and reliable. The enzyme activity and glycogen content of the ADP-glucose pyrophosphorylase mutant are evidently higher than those of wild corn AGPase, evident change of the substrate affinity of the ADP-glucose pyrophosphorylase mutant is avoided, and the sensitivity of the ADP-glucose pyrophosphorylase mutant to activating agent is increased.