50 results about "18s rdna" patented technology

The invention provides a liriope spicate endogenous aspergillus fungus and application thereof to preparation of steroid saponin, and relates to the technical field of microorganisms. The fungus mainly solves the problems that due to the factors that the liriope spicate steroid saponin is mostly and directly from liriope spicate extracts, the liriope spicate steroid saponin content is low, the saponin extraction cost is high, and the like, the market delivery quantity of the liriope spicate steroid saponin is limited. The aspergillus fungus is separated from living body tuberous roots of liriope spicata var. prolifera, and belongs to aspergillus sp. through 18S rDNA sequence identification; the aspergillus fungus is preserved in CCTCC (China Center for Type Culture Collection); the preservation data is December 7, 2016; the preservation number is CCTCC No.M2016721. The liriope spicate produced by a liriope spicate GAP (Good Agricultural Practices) base (the liriope spicate source areain Oumao town, Xiangyang city in Hubei province ) is used; the liriope spicate steroid saponin is generated through liriope spicate endogenous fungus plant liquid fermentation. The endogenous fungus is an important microorganism for finding a new resource of the liriope spicate steroid saponin.

Tools and methods for detecting the presence bacteria, yeast and mold in a sample obtained from a food sample are provided. The methods employ a polymerase chain reaction and primer and probe sets that are based on the 16S rRNA and squalene-hopene cyclase genes of Alicyclobacillus and Geobacillus and the 18S rDNA gene of mold and yeast. The present invention also relates to primer and probe sets. Each primer and probe set comprises a forward primer and a reverse primer, both of which are from 15 to 35 nucleotides in length and a probe.

The invention relates to an analysis method for plankton type community DNA dactylogram in urban sewage, the method is as follows: metagenome of plankton type community in urban sewage is amplified through using a primer aiming at 16S rDNA and 18S rDNA, and undergoes denaturing gradient gel electrophoresis to obtain a fingerprinting of community DNA. Firstly, the metagenome DNA of the plankton type community is extracted; secondly, the object gershgorim band of the metagenome is amplified; thirdly, the DGGE electrophoretic separation is performed to obtain the fingerprinting of the plankton type community DNA; fourthly, a DGGE gel picture is analyzed through using DNA UVI software, using the optical density value of the gershgorim band to indicate the gershgorim band; and finally, data analysis is performed through using statistics software. The analysis method for plankton type community DNA dactylogram in urban sewage has the advantages of convenience and efficiency, excellent repeatability, high sensitivity and low cost, capability of being applied to biological monitoring of urban sewage.

The invention discloses an expression carrier of an industrial microorganism that is candida utili and simultaneously provides a construction method of the candida utili expression carrier. The invention uses a basic skeleton of a pBR322 carrier and a template of candida utili genomic DNA, obtains a cycloheximide resistance gene with the site-directed mutagenesis method, obtains a rDNA segment and a GAP promoter-terminator segment of the candida utili genomic DNA by adopting the PCR method and the template of the candida utili genomic DNA, and constructs the integrated candida utili expression carrier that is mediated by 18S rDNA and has cycloheximide resistance. The integrated candida utili expression carrier can be used for preparing a foreign candida utili expression protein and can be widely applied into the fields of feedstuff addition agents, food industry, and pharmacy, and the like.

The invention relates to scenedesmus capable of highly yielding oil as well as a culture method and application thereof. An alga variety separated from a eutrophic water region is subjected to 18S rDNA sequence analysis and microstructure morphology identification so that the alga is determined as the member of Scenedesmus sp. and the sequence number is KF999043. The alga strain is named as Scenedesmus quadricauda SDEC-8 and is preserved in China Center for Type Culture Collection on October 8, 2014; and the preservation number is CCTCC NO: M2014448. An oil production culture method comprises the following steps: separating single alga cells of the Scenedesmus quadricauda SDEC-8 and carrying out aeration enlarged cultivation on obtained pure cells under laboratory conditions. The Scenedesmus quadricauda SDEC-8 contains rich high-quality oil; and the oil content can be up to 31.8% of the dry weight of the cells and the content of saturated fatty acids in fatty acids can be up to 80%, so that the Scenedesmus quadricauda SDEC-8 can be used as the raw material of biodiesel. Based on a high cetane number (61.38) and a low iodine number (29.38gI2 / 100g), the biodiesel prepared by taking the microalga as the raw material has good oxidization stability.

The invention relates to a constructing method of a standard sample in Alexandrium catenella (A.catenella) fluorescence quantitative polymerase chain reaction. A recombinant plasmid is constructed by extracting DNA of A.catenella ACDH01, amplifying 18S rDNA of A.catenella ACDH01 to a 28S rDNA gene segment through polymerase chain reaction, and connecting an amplified product with a T carrier; the confirmed recombinant plasmid is diluted into a polymerase chain reaction standard sample in a gradient way; real-time polymerase chain reaction is carried out by using 5.8S-b5' and 5.8S-b3' as primers and a recombinant plasmid standard sample as a template; and a standard curve is prepared according to a clinical circulating number in the reaction and the log value of the gradient concentration of the recombinant plasmid standard sample. The recombinant plasmid real-time polymerase chain reaction standard sample with 18S rDNA-28S rDNA gene segments of the A.catenella ACDH01 is established; and the real-time polymerase chain reaction detection is accurately, efficiently and rapidly carried out on A.catenella ACDH01 to be tested by adopting a pair of universal primers.

The invention discloses a rhodosporidium sphaercarpum ZDFY1801 and application thereof. The preservation number of the strain is GDMCC No: 60679. The strain is identified as rhodosporidium sphaercarpum through a 18S rDNA gene.The strain ZDFY1801 has the capability of efficiently removing ammonia nitrogen in simulated aquaculture water, and also has flocculation and clarification effects on organicsuspended particles in the aquaculture water. The strain ZDFY1801 can also significantly inhibit vibrio of aquaculture water, and the strain ZDFY1801 has high biological safety and also has rich nutrition. The characteristics show that the ZDFY1801 can be used for treating wastewater in aquaculture and can also be used as bait for aquaculture animals or used as a feed additive.

Oligonucleotide molecules for the detection of Giardia lamblia (G. lamblia which molecules hybridise under medium to high stringency conditions to unique 18S rDNA / rRNA sequences of G. lamblia, and methods for the detection of the presence of viable cells of G. lamblia in samples using the oligonucleotide molecules.

The invention discloses a method for analyzing plant endophyte colonies. In the method, a primer pair used for analyzing the plant endophyte colonies is provided, the target sequence of the primer pair is the overall length or part of the region from V3 to V4 of bacteria 16SrDNA; there are differences between the last basic group at the 3' end of one primer in the primer pair and the plant mitochondria 18S rDNA, and the number of the difference sites between the overall length of the primer and the plant mitochondria 18S rDNA is equal to or larger than 3; there are differences between the lastbasic group at the 3' end of the other primer and the plant chloroplast 16S rDNA, and the number of the difference sites between the overall length of the primer and the plant mitochondria 18S rDNA is equal to or larger than 3. The two primers 322F-1 and 796R are designed, the DNA polymerase with hot-start activity but without 3' to 5' exonuclease proofreading activity is selected to achieve theaim that the pollution of the host plant chloroplast 16S rDNA and the pollution of the host plant mitochondria 18S rDNA are avoided at the same time, and therefore, a pure bacteria colony 16S ampliconlibrary is obtained.

The invention provides a DGGE / TGGE (Denaturing Gradient Gel Electrophoresis / Temperature Gradient Gel Electrophoresis) analysis method of specific 18S rDNA (Deoxyribose Nucleic Acid) fragment without being based on a GC clamp strategy. The specific 18S rDNA fragment is a fragment containing a terminal region of 18S rDNA, which is far away from an ITS (Internal Transcript Space) sequence; and with a 18S rDNA sequence of a Saccharomyces cerevisiae standard strain as a reference, the terminal region of the 18S rDNA, which is far away from the ITS sequence, is equivalent to a fragment which starts from any nucleotide within a range of the first to the 30th and ends with the 291th-locus nucleotide. According to the invention, a result consistent with that obtained by the use of the GC clamp can be obtained under the same condition by adopting the DGGE / TGGE analysis of the specific 18S rDNA without adopting the GC clamp, therefore, the experiment cost can be reduced and the experiment operation steps are reduced.

The invention discloses a pleuronectiformes soleidae fish ribosome ITS1 (internal transcribed spacer region 1) universal primer design method and application. The method has the beneficial effects that the disclosed primer design method is designed on the basis of the conservative 18S rDNA and 5.8 S rDNA sequences at the two ends of ribosome ITS1; the obtained primer has very high specificity; theamplified ITS1 evolutionary rate is relatively high; similarity basically shows within the variety; the obvious differences are shown between varieties; when the primer is applied to soleidae fishes,the accuracy is high; the form feature identification defects are overcome; the problem of idioplasm mixing is solved.

The invention relates to the technical field of microorganisms, in particular to laetiporus cinnabarinus and application thereof. The strain provided by the invention has a 18S rDNA-ITS sequence which is at least 99% consistent with the sequence as shown in SEQ ID NO: 1. The strain provided by the invention can significantly reduce the level of interleukin 6 (IL-6) and tumor necrosis factor (TNF-alpha) so as to treat and prevent interleukin 6 (IL-6) and / or tumor necrosis factor (TNF-alpha) pathway abnormality related diseases, and has the advantages of fast effect, greenness, safety and reliability.

The invention discloses a method for obtaining desmodesmus NMX451 and a genetic transformation method of the desmodesmus NMX451 and relates to screening of oil-producing microalgae and a genetic transformation technology thereof. Green alga separated from a wild environment is subjected to 18S rDNA (ribosomal deoxyribonucleic acid) sequence analysis and morphological identification and is determined to be a desmodesmus member, and a sequence of the green alga is SEQ NO.1. The genetic transformation method of the green alga comprises the following steps: (1) constructing a nuclear gene transformation plasmid with a resistance gene; (2) transforming the desmodesmus NMX451 by adopting an electrotransfer method; (3) detecting whether a transformant is positive or nor through PCR (polymerase chain reaction); (4) proving that an exogenous gene enters host DNA (deoxyribonucleic acid) through DNA blot hybridization. The desmodesmus NMX451 can tolerate high-concentration sodium bicarbonate, and oil content can be 27.63% of dry cell weight; algae cells of the desmodesmus NMX451 also contain high proportion of linolenic acid; by adopting the method for obtaining the desmodesmus NMX451, the exogenous gene can be introduced into the desmodesmus NMX451, so that the desmodesmus NMX451 can obtain resistance to zeocin and also can express a protein with an economic value.

The invention discloses a specific gene of brown atrichopogon and a molecular identification method of the brown atrichopogon. The specific gene of brown atrichopogon and the molecular identification method of the brown atrichopogon are characterized in that a molecular biology method is adopted, the gene sequence 18S rDNA of the atrichopogon is obtained, and through comparison of gene sequence similarity, species identification is conducted on the atrichopogon. By means of the molecular identification method of the brown atrichopogon, it is facilitated to achieve accurate and rapid identification of the brown atrichopogon.

The invention discloses a taxol-producing endophytic fungus and relates to taxol-producing fungi. The invention provides a new taxol-producing endophytic fungus and increases the number of strains that can be used in microbial fermentation of taxol. The taxol-producing endophytic fungus HD86-9 belongs to aspergillus niger yew variants and is preserved in China Center for Type Culture Collection with the preservation date of December 25th, 2006 and the preservation number of CCTCC M 206137. The taxol-producing endophytic fungus HD86-9 belongs to a new aspergillus niger yew variant under the analysis of morphology, 18S rDNA sequence and ITS. The invention not only provides an additional taxol-producing endophytic fungus, but also establishes physical foundation for the genetic modification of taxol-producing microorganisms in future.

The invention discloses Xiangyang radix ophiopogonis endophytic Fusarium sp. fungus and application in steroid saponin preparation, and relates to the technical field of microorganisms. The problems that Xiangyang radix ophiopogonis steroid saponin is mostly directly from Xiangyang radix ophiopogonis extractives, and the Xiangyang radix ophiopogonis steroid saponin is limited in market delivery quantity due to the fact that Xiangyang radix ophiopogonis is long in earthnut production period, low in steroid saponin content and high in saponin extraction cost are solved. The endophytic fungus isseparated from Xiangyang radix ophiopogonis (Liriope spicata var.proliferaY.T.Ma) living earthnuts, it is determined as Fusarium sp. through 18S rDNA sequence authentication, the penicillium oxalicumis preserved in the China Center for Type Culture Collection, the preservation date is 7th December, 2016, and the preservation number is CCTCC No:M2016723. By means of Xiangyang radix ophiopogonis produced in a Xiangyang radix ophiopogonis GAP base-the source area of Xiangyang radix ophiopogonis in Oumiao county, Xiangyang city, Hubei province, due to fermentation of Xiangyang radix ophiopogonisendophytic fungi strain liquid, the Xiangyang radix ophiopogonis steroid saponin is produced. The endophytic fungus is the important microorganism for searching for new resources of the Xiangyang radix ophiopogonis steroid saponin.

The invention provides an expression vector of schizochytrium limacinum as well as a construction method and application of the expression vector, and belongs to the field of gene engineering. The expression vector comprises a pSc-18S rDNA (ribosomal deoxyribonucleic acid) vector, a combination of a promoter and a terminator of schizochytrium limacinum, and a gene sequence of a resistance-fluorescence double selection marker, the combination of the promoter and the terminator comprises at least one of the following combinations: a combination of a promoter PT and a terminator TT, a combination of a promoter PA1 and a terminator TA, a combination of a promoter PA2 and a terminator TA, and a combination of a promoter PE2 and a terminator TE. According to the present invention, with the efficient schizochytrium limacinum genome editing and transformant screening technology, the resistance-fluorescence double-selection marker is efficiently integrated to the schizochytrium limacinum chromosome through the gene gun in the fusion protein expression form, such that the gene editing element test and the transformant rapid screening can be performed based on the expressed fluorescence intensity; especially, when endogenous genes are expressed, the technical advantages are very prominent.

The invention discloses a creat endophytic fungus strain LM19J01 and application thereof. The strain LM19J01 belongs to fusarium in taxology, is named Fusarium sp., and was collected by China Center for Type Culture Collection on November 11, 2015; and the collection number is CCTCC NO:M2015659. The genome 18s rDNA (ribosomal deoxyribonucleic acid) base sequence of the strain is disclosed as SEQ ID NO.1. The experiment proves that the creat endophytic fungus strain LM19J01 can perform biotransformation on neoandrographolide to obtain the transformed compounds.

The invention discloses Xiangyang radix ophiopogonis endophytic penicillium oxalicum and application in steroid saponin preparation, and relates to the technical field of microorganisms. The problemsthat Xiangyang radix ophiopogonis steroid saponin is mostly directly from Xiangyang radix ophiopogonis extractives, and the Xiangyang radix ophiopogonis steroid saponin is limited in market delivery quantity due to the fact that Xiangyang radix ophiopogonis is long in earthnut production period, low in steroid saponin content and high in saponin extraction cost are solved. Penicillium oxalicum isseparated from Xiangyang radix ophiopogonis (Liriope spicata var.proliferaY.T.Ma) living earthnuts, it is determined as penicillium oxalicum through 18S rDNA sequence authentication, the penicillium oxalicum is preserved in the China Center for Type Culture Collection, the preservation date is 7th December, 2016, and the preservation number is CCTCC No:M2016722. By means of Xiangyang radix ophiopogonis produced in a Xiangyang radix ophiopogonis GAP base-the source area of Xiangyang radix ophiopogonis in Oumiao county, Xiangyang city, Hubei province, due to fermentation of Xiangyang radix ophiopogonis endophytic fungi strain liquid, the Xiangyang radix ophiopogonis steroid saponin is produced. The endophytic fungi are the important microorganism for searching for new resources of the Xiangyang radix ophiopogonis steroid saponin.

The invention provides a candida utilis expression vector for producing laccase, a construction method of the candida utilis expression vector, recombinant engineering bacteria and an application of the recombinant engineering bacteria. The candida utilis expression vector for producing the laccase comprises a 18S rDNA gene segment, a yeast glyceraldehyde triphosphate dehydrogenase gene promoter sequence (GAP-p), a laccase gene, a yeast glyceraldehyde triphosphate dehydrogenase gene terminator sequence (GAP-t) and a cycloheximide (CYH) resistance gene. According to the invention, all genetic elements of the whole expression system are from probiotic candida utilis and non-toxic and harmless fungi, antibiotic resistance genes are not carried, drifting, diffusion and integration of the resistance genes do not exist, other toxic protein genes are not carried, biosafety hazards to the environment or people and animals are avoided, the food grade is achieved, and the recombinant engineeringbacteria can be directly added into foods, medicines and feeds for application.

The invention discloses a primer for inhibiting sequence-specific amplification of 18S rDNA of shrimps and oysters. The primer mainly comprises base sequences as shown in SEQ ID NO.1, five deoxyhypoxanthines inserted among the base sequences AG, and C3 spacers at the tail ends of the base sequences. The primer can effectively inhibit the amplification of host genes and enhance the amplification effect of 18S rDNA of other eukaryotes (such as protozoa, algae and fungi) in host bodies. The invention further discloses the application of the primer in preparing a reagent with the effect of inhibiting the sequence-specific amplification of 18S rDNA of the shrimps and oysters.

The invention provides a liriope spicate endogenous schizophyllum fungus and application thereof to preparation of steroid saponin, and relates to the technical field of microorganisms. The fungus mainly solves the problems that due to the factors that the liriope spicate steroid saponin is mostly and directly from liriope spicate extracts, the liriope spicate steroid saponin content is low, the saponin extraction cost is high, and the like, the market delivery quantity of the liriope spicate steroid saponin is limited. The schizophyllum fungus is separated from living body tuberous roots of liriope spicata var. prolifera, and belongs to schizophyllum sp. through 18S rDNA sequence identification; the schizophyllum fungus is preserved in CCTCC (China Center for Type Culture Collection); thepreservation date is December 20th, 2016; the preservation number is CCTCC No. M2016765. The liriope spicate produced by a liriope spicate GAP (Good Agricultural Practices) base (the liriope spicatesource area in Oumao town, Xiangyang city in Hubei province ) is used; the liriope spicate steroid saponin is generated through liriope spicate endogenous fungus plant liquid fermentation. The endogenous fungus is an important microorganism for finding a new resource of the liriope spicate steroid saponin.

The invention provides a method for evaluating the diversity of marine zooplankton on the basis of a macro bar code technology, belonging to the field of marine organism diversity research. The methodcomprises the following steps: DNA extraction of a sample metagenome, PCR amplification of a V4 hypervariable region fragment of 18S rDNA to construct a library, high-throughput sequencing, and bioinformatic analysis, wherein a PCR primer used for library construction comprises a specific tag sequence. The method provided by the invention is simple and convenient to operate, can improve the output of sequencing sequences, increases the resolution rate of a sequence with a tag, can improve the amplification efficiency of a template with high GC content, reduces the generation of repetitive sequences, improves the actual expression quantity obtained by sequencing, and increases the accuracy of evaluating the diversity of the marine zooplankton.

The invention discloses widmannella terrestris with high yield of EPA grease and an application of the widmannella terrestris. Algal strain WL1 separated from desert organism soil crust is subjected to 18S rDNA sequence analysis and microscopic morphology identification to determine that the algal strain WL1 belongs to Widmannella. According to the algal strain, in a batch culture mode of the algal strain, neutral fat accumulated in algal cells accounts for more than 80% of total fat, fatty acid mainly comprises palmitoleic acid, oleic acid and linolenic acid and can account for more than 70% of total fat fatty acid, the algal strain is suitable for production of biodiesel, contains unsaturated fatty acid eicosapentaenoic acid required by a human body, and the content of the fatty acid accounts for more than 10% of the total fatty acid.

The invention discloses an endogenous penicillium fungus of radix liriopis proliferae and application thereof in preparation of steroidal saponin, and relates to the technical field of microorganisms.The endogenous penicillium fungus of the radix liriopis proliferae and the application thereof in preparation of the steroidal saponin mainly solve a problem that the steroid saponin of the radix liriopis proliferae is limited in market due to factors of long tuberous production cycle, low steroidal saponin content and high extraction cost of saponin of the radix liriopis proliferae. The endogenous penicillium fungus is isolated from living roots of the radix liriopis proliferae (Liriope spicata var. prolifera). The endogenous penicillium fungus of the radix liriopis proliferae is identifiedby 18S rDNA sequence as Penicillium sp and is deposited with China Center for Type Culture Collection (CCTCC) on December 7, 2016 with the preservation number of CCTCC No. M2016724. According to the invention, the radix liriopis proliferae produced in radix liriopis proliferae source area, Oumiao town, Xiangyang city, Hubei Province-- radix liriopis proliferate GAP base is used, radix liriopis proliferae steroidal saponin is produced through liquid fermentation of endogenous penicillium fungus strains of the radix liriopis proliferae. The endogenous penicillium fungus is an important microorganism for searching for new resources of the radix liriopis proliferae steroidal saponin.

The invention provides a temperature-sensitive bacterial strain for producing polyunsaturated fatty acids, the preservation number is CGMCC 11808, and through 18S rDNA identification, the bacterial strain is a temperature-sensitive filamentous fungus of the genus Mortierella. The growth temperature is 25°C, and the temperature sensitive range is 25°C-28°C. After 18S rDNA identification, the strain is Mortierella alpina. The strain can grow well at 20°C-25°C. When the temperature is higher than At 26°C, the hyphae cannot extend normally. When the temperature is higher than 27°C, the bacterial colony becomes significantly smaller; when the temperature is higher than 30°C, the bacteria do not grow at all. Under the condition of 25°C, the strain was fermented with fermentation medium, and it was found that it could produce various unsaturated fatty acids.

The invention discloses a PCR specific primer and a detection method for detecting Schizospora anguilla. The sequence of an upstream primer of the primer pair is shown as SEQ ID No.1, and the sequence of a downstream primer is shown as SEQ ID No.2. DNA of a sample to be detected is extracted, PCR amplification is carried out by taking the DNA as a template and taking the primer pairs as shown in SEQ ID No.1 and SEQ ID No.2 as primers, and if an 836bp amplification product can be obtained, the sample contains the Schelyriasis anguilla. The primer is designed by using the Schizospora anguilla 18S rDNA as a molecular target, the Schizospora anguilla can be successfully expanded from eel and siniperca chuatsi tissues infected with the Schizospora anguilla, and the primer has high specificity and sensitivity, is simple and rapid to operate, can accurately detect and identify the early stage of the Schizospora anguilla, and the eel skin sporidiosis can be effectively predicted.