369 results about "MOLECULAR BIOLOGY METHODS" patented technology

The invention discloses a method and an automatic instrument device for enriching and extracting target cells and separating single cells by using a positive magnetic bead method and performing immunity and molecular biology identification and analysis on single cells. By the method and the instrument device, the on/off of a capture magnet and a release magnet is controlled by various methods to complete target cell searching, capturing, cleaning and releasing operation once or for multiple times, so that the target cell detection sensitivity and stability are improved. When the capture magnet searches and captures the target cells, the search line is circular, square, comb-shaped, S-shaped or U-shaped. In addition, the captured substances are filtered, most free micro magnetic beads are removed, the purity of the product is further improved and the product can be used as a good experimental material. By combining a special filter and the adsorption of the capture magnet, more than 95 percent of free micro magnetic beads can be effectively removed. Meanwhile, the types of the single cells are identified and biological characteristics of the single cells are analyzed by an immunofluorescence staining method and a method in molecular biology, and effective biological indexes are provided for clinical diagnosis and treatment of cancers.

A method of using elongate multicellular organisms in conjunction with a specialized flow cytometer for drug discovery and compound screening. A stable, optically detectable linear marker pattern on each organism is used to construct a longitudinal map of each organism as it passes through the analysis region of the flow cytometer. This pattern is used to limit complex data analysis to particular regions of each organism thereby simplifying and speeding analysis. The longitudinal marker pattern can be used to alter signal detection modes at known regions of the organism to enhance sensitivity and overall detection effectiveness. A repeating pattern can also be used to add a synchronous element to data analysis. The marker patterns are established using known methods of molecular biology to express various indicator molecules. Inherent features of the organism can be rendered detectable to serve as marker patterns.

The invention discloses a method for adjusting and controlling microbial enhanced oil recovery, which comprises the following steps: (1) analyzing the microbial community structure in produced fluid of an oil reservoir via molecular biological method and/or detecting the metabolites in the produced fluid; (2) adjusting the microorganism(s) to be injected into the oil reservoir and/or the nutrient system corresponding to the microorganism(s); (3) injecting the adjusted microorganism(s) and/or the nutrient system corresponding to the microorganism(s) into the oil reservoir through a water injection well; and (4) obtaining the crude oil from a corresponding beneficial oil producing well. Compared with the prior art, the method of the present invention adjusts microbial community structure in the oil reservoir to evolve toward the direction of facilitating oil production, and the performance of the functional microorganism(s) can be completely realized; the nutrient system is pertinently injected to avoid the blindness of using the nutrient system. Therefore, the method is scientific, economical and effective for the microbial enhanced oil recovery.

The invention relates to an RNA-dependent RNA-polymerase, to methods and kits for marking and/or amplifying in a primary dependent or independent manner a ribonucleic acid (RNA), in particular a viral, eucaryontic, procaryontic and double-stranded ribonucleic acid (RNA) for the biological and medical use thereof. Said RNA-dependent RNA-polymerase comprises a right-hand conformation and the amino acid sequence the following sequence sections: a. XXDYS b. GXPSG c. YGDD d. XXYGL e. XXXXFLXRXX having the following significances: D: aspartate, Y: tyrosine, S: serine, G: glycine, P: proline, L: leucine, F: phenylalanine, R: arginine, X: any amino acid. The inventive amplifying method is particularly suitable for microarray engineering, siRNA production and for diagnosticating viral infections by detecting viral RNA in patient samples. The inventive marking method is particularly suitable for purifying RNA by means of affinity bonds and for marking RNA used in molecular biology methods for characterising the function and/or structure of a viral, eucaryontic, procaryontic and double-stranded ribonucleic acid (RNA).

The present invention relates to DNA polymerases with a special mutation which have an enhanced mismatch discrimination, the preparation and use thereof. The thermostable DNA polymerases with this mutation are particularly suitable for diagnostic and molecular-biological methods, e.g., allele-specific PCR.

The invention relates to a molecular biology method for identifying purity of tobacco varieties. The method comprises the following main steps: respectively extracting total genome DNA of a control group and a to-be-detected tobacco variety, performing whole genome sequencing of proper depth by adopting the latest sequencing technology, performing sequence splicing, assembling and whole genome sequence comparison on the control group and the to-be-detected tobacco variety based on a tobacco genome reference sequence by utilizing bioinformatics means, counting base differences of the two groups, and calculating the purity percentage of the to-be-detected tobacco variety relative to the control tobacco variety. The method disclosed by the invention is not influenced by environmental conditions and seasons, is accurate and reliable in result, can accurately identify the purity of the tobacco varieties from a single base variation level of the smallest hereditary unit, can be used for parent purification and authenticity identification of the tobacco varieties and has great significances for guaranteeing safety of tobacco production varieties, maintaining economic benefits of tobacco growers and effectively solving intellectual property disputes of the tobacco varieties.

A use of PD-0332991 in preparation of drugs preventing and treating drug-resistant tumor is disclosed. Application of a cell biological method and a molecular biological method and human gefitinib-resistant nude-mouse transplanted tumor model tests prove that: the PD-0332991 can effectively inhibit growth of human gefitinib-resistant lung cancer cells, induce apoptosis, regulate and control cell cycle related gene, cause tumor cell cycle arrest and induce apoptosis. Accordingly, the PD-0332991 can be adopted as an effective component for preparation of drugs preventing and treating gefitinib-resistant tumor, and foods, health products and cosmetics for preventing and treating drug-resistant tumor.

The related rice gene mark for broad spectrum anti rice blast includes mark length 10-30 nucleotides with sequence same as the sequence of 1st-70798th in SEQ ID No:1. Wherein, based on our rice resource and natural or induced diseases, it applies different rice blast plants with the domestic as main and foreign as assist to screen the 'Gumei 4th', uses molecular mark technique to locate gene and obtain Pi-gm(t) gene with chain mark, and has great application value in breeding.

The invention discloses a method for quickly identifying Heliothis armigera and Helicoverpa assulta, and belongs to the field of molecular biology. The method comprises the following steps of: extracting total DNA by adopting a standard phenol-chloroform protocol or a DNA extraction kit; amplifying a front sequence of mitochondria CO1 by using a DNA barcode primer, and using the amplified sequence as a mark; and performing diagnostic characters of nucleotides. By the method, different biotypes and larvae or incomplete individuals of the same variety of Heliothis armigera and Helicoverpa assulta can be identified; and the method has the characteristics of quickness and convenience, and is more accurate and reliable than the traditional morphologic identification.

The invention belongs to the technical field of rape breeding, and discloses a selection breeding method for a Brassica napus-turnip cabbage cytoplasmic male sterility line (CMS). The method is characterized in that an artificially-bred turnip cabbage allotetraploid as a female parent is hybridized with a Brassica napus variety as a male parent, male sterile plants are isolated from the progeny of hybrid selfing, and a Brassica napus-turnip cabbage cytoplasmic male sterility line with genetic stability is obtained by successive backcross breeding. Restorer line and maintainer line relationship and molecular biological methods are employed to prove genetically and on DNA level respectively that the Brassica napus-turnip cabbage cytoplasmic male sterility line has a sterile cytoplasmic type different from those of Polima cytoplasmic male sterility (pol CMS), turnip cytoplasmic male sterility (ogu CMS, kos CMS) and Brassica juncea cytoplasmic male sterility (hau CMS). The invention adds a new rape cytoplasmic male sterility type for China and the rest of the world.

The invention relates to a novel gene sequence of a recombinant chicken Alpha interferon, constructs a recombinant expression vector thereof and belongs to a gene engineering biological product obtained by a molecular biology method. The gene sequence of a newly designed chicken Alpha interferon is recombined into a pPICZ Alpha-A vector and then is confirmed on the special position of a microzyme by an electric conversion mode. Besides, a pichia expression system is adopted to express a foreign gene, thus being beneficial to the commercial production of the chicken interferon. The protein expressed by a gene group after being diluted by 4365158.3 times can completely restrain the attraction of vesicular stomatitis virus of 100-1000TCID50. Compared with a natural chicken Alpha interferon, the novel gene sequence of a recombinant chicken Alpha interferon has a higher anti-virus effect; the anti-virus effect thereof is improved by 8 times. Test also detects that the protein expressed by the gene can restrain the proliferation of a newcastle disease virus and an avian influenza virus; besides, the effect of the interferon with high concentration is more remarkable.

The present invention discloses a performance improved recombination staphylococcus aureus protein A affinity ligand and a construction method thereof. The present invention adopts a molecular biology method. A sequence B of a nature protein A is selected for molecular transformation. A C-terminal of the sequence B is added with two cysteines, so that the protein A can pass through double-locus coupled chromatography matrix to stabilize the connection. Six glycines are added to the end of a second Loop of the sequence B to increase the length and reduce the binding force with an antibody, so that elution conditions are mild. On this basis, resistance performance to high concentration base of the protein A is transformed. Asparagines and phenylalanine at 23rd and 30th positions of the sequence B are respectively replaced by threonine and alanine to obtain a sequence Z with higher alkaline resistance properties. Then, isocaudarner is used for connecting sequence Zs of different numbers head-to-tail in series. The efficient expression system of e. coli is used for overexpression. The expressed recombination protein A is coupled to agarose matrix preparation affinity chromatography fillers and is used for purifying antibodies. Results show that the recombination protein A affinity ligand prepared by the present invention is good in elution performance and alkali resistance.

The invention relates to a phospholipase B from pseudomonas fluorescens and a production method thereof, which belong to the field of enzyme gene engineering and enzyme engineering. The phospholipase B is produced from the pseudomonas fluorescens, has the total gene length of 1272bp and codes of 423 amino acids, and the zymoprotein theoretical molecular weight is 45.8kDa, wherein 1 to 69bp code phospholipase B signal peptide, and 70 to 1272bp code phospholipase B mature peptide are comprised. An expression vector and a recombinant host of the phospholipase B can be obtained by the traditional molecular biological method, i.e. colibacillus recombinant plasmid and recombinant colibacillus containing genes of the phospholipase B are obtained. The phospholipase B has good activity and stability at low temperature, does not have the lipase activity, can be hydrolyzed for catalyzing two fatty acyl group radicals in complete phospholipids, has wide application in the industrial process of grease refining, phospholipids emulsifying agent modification and the like, and also has simple preparation method, high product quality and low production cost.

The invention relates to an RH blood type DEL-type RHD93T>A allele and a detection method thereof, and belongs to the technical field of molecular biology analysis and detection. According to the detection method, primer sequences capable of amplifying areas including target sites with mutant gene sequences are designed through a PCR-specific primer technology; PCR amplification is selectively conducted on the areas of the target sites including the mutant genes through a gene amplification method so as to specially detect specific RHD93T>A allele. The detection method adopts a molecular biology method for gene-level detection, so that the detection sensitivity and specificity are improved, and the detection method is simple, convenient and fast. Due to differences in RHD gene of different ethnic groups, the detection method is designed specially for RHD93T>A allele on the basis of correlational research and according to the Chinese RHD gene molecule background. The RH blood type DEL-type RHD93T>A allele and the detection method thereof not only are of important clinical practical significance in making up for deficiencies of the serological technique, but also has a wide application value on scientific research.

The invention discloses a molecular-biological method for quickly distinguishing prodenia litura and other two noctuidae pests (cotton bollworm and tobacco budworm). The method comprises the following steps of: extracting total DNA (Deoxyribonucleic Acid) by adopting a standard phenol-chloroform method or a DNA extraction kit; using a DNA bar code primer to amplify a sequence at the front part of mitochondria CO1 as a label; and comparing diagnostic characters of nucleotides. The molecular-biological method can be used for distinguishing different biotypes and larvae or incomplete individuals of the prodenia litura, the cotton bollworm and the tobacco budworm, has the characteristics of accuracy, quickness, convenience and quickness, and is more accurate and reliable compared with the traditional morphological identification.

The invention discloses primers, a kit and a detection method for detecting a gene type of a dominant white feather site of chicken PMEL17 gene and belongs to the technical field of biology. According to the detection method, a specific primer P1-R of allelic gene I is designed, aiming at a situation that a 9bp insert (-CTGGGCACC-) exists in a 10th exon of the PMEL17 gene; a specific primer P2-F of allelic gene i is designed, aiming at the situation that no 9bp insert exists in the 10th exon of the PMEL17 gene; the specific primer pair P1 of the allelic gene I is used for amplifying an allelic gene I of the PMEL17 gene; the specific primer pair P2 of the allelic gene i is used for amplifying an allelic gene i of the PMEL17 gene; after a PCR (Polymerase Chain Reaction), agarose gel electrophoresis is carried out; the gene type of the dominant white feather site of the PMEL17 gene is detected accurately, rapidly and conveniently according to the result of the agarose gel electrophoresis. Compared with the prior art, a molecular biological method established in the invention can greatly improve the judging efficiency and accuracy of the gene type; and the method is simple, easy to operate and low in cost.

The present invention relates to human papillomavirus type bivalent virus-like particle mixture, and human papillomavirus type bivalent virus-like particle mixing protein antigen and its construction method. The present invention includes one kind of cervical carcinoma virus HPV16 L1 antigen and one kind of condyloma acuminata virus HPV11 L1 antigen. The HPV11 L1 gene is first cloned from condyloma acuminata tissue through molecular geological method, and both HPV16 L1 gene cloned from cervical carcinoma tissue and the HPV11 L1 gene are then expressed in the insect cell expression system of baculovirus, so as to constitute recombinant virus strain Bacmid/HPV16-HPV11L1. Animal immunizing test shows that the constituted recombinant protein has excellent immunogenicity and immunoreactivity, and may be used as candidate antigen of gene engineering multivalent vaccine for preventing condyloma acuminata and cervical carcinoma.

The present invention discloses a method that can be used to identify one DNA sequence or one specific group of DNA sequences from a complex biological sample. Diverse molecular biology methods require the use of short DNA sequences, called oligonucleotides, that are artificially synthesized from a description of their composing bases. The disclosed method allows the design of oligonucleotides useful for said molecular biology procedures, like probe design procedures, and is characterized by the construction of a database of reference sequences, the selection of a subset of sequences belonging to target organisms, the selection of candidate oligonucleotides from such sequences, the depuration of these candidate oligonucleotides according to hybridization specificity and thermodynamic stability criteria, and the sorting of such oligonucleotides according to their taxonomic specificity. In a second aspect, a method is disclosed to design oligonucleotides pairs or primers, which are required in certain molecular biology techniques, like polymerase chain reaction (PCR) techniques. This method is similar to the first aspect of the invention, but thermodynamically compatible oligonucleotides pairs or primers that hybridize to the same sequence at a distance which is within a given range are evaluated.

The invention discloses application of isothiocyanate to the preparation of medicine for preventing and treating medicine-resistance tumor. The invention uses a celluar and molecular biology method for proving that the isothiocyanate can effectively inhabit the growth of medicine-resistance tumor cells, induce the apoptosis, regulate and control the tumor medicine-resistance relevant genes, cause medicine-resistance tumor cell cycle arrest, and cause tumor cell oxidative damage. Thereby, the isothiocyanate can be used as a active ingredient for preparing the medicine for preventing and treating medicine-resistance tumor, and food, health-care products and cosmetics for preventing and treating medicine-resistance tumor.

The invention belongs to the field of molecular biology method and relates to a reagent composition for separating total RNA in a plant or a microorganism and a preparation method thereof. The high quality RNA can be obtained after a simple extraction by chloroform while guanidinium isothiocyanate and phenol are regarded as main components, positive ions are provided by NaCl, MgCl2 and the like, and a solution pH is stabilized by a sodium acetate-acetic acid buffer system. Glycogen serving as a nucleic acid precipitant is added in the RNA extraction reagent in the invention, so that the reagent disclosed by the invention, in comparison with reagents of the same type, greatly improves precipitation efficiency of the nucleic acid and also can efficiently separate the nucleic acid component which has a very low content in a tissue sample. Therefore, the nucleic acid precipitation process can be finished in a short period, the precipitation process at -20 DEG C for hours is avoided, and operating time is shortened greatly. The method is rapid as well as efficient and has wide applicable samples; and the reagent composition disclosed by the invention is cheaper than commercial TRIzol reagents. The disadvantages of high sample selectivity, fussy operation and relatively low efficiency of the traditional method are overcome. The operation is more flexible; and the sample after being homogenized can be stored at -20 DEG C and then used for the RNA extraction.