49 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 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 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 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 liriope spicatavar endogenous aspergillus niger and application thereof in preparing steroid saponin, relates to the technical field of microorganisms and mainly solves the problem that liriope spicatavar steroid saponin market release quantity is limited caused by the fact that liriope spicatavar is long in tuberous root production period, low in steroid saponin content and high in saponin extraction cost because most liriope spicatavar steroid saponin directly comes from liriope spicatavar extract at present. Endogenous fungus is separated from living tuberous root ofLiriope spicatavar. Prolifera Y. T. Ma, identified as Aspergillus niger through 18S rDNA sequence and already collected at the China center for Type Culture Collection on Dec. 15, 2016 under the number of CCTCC No:M2016753. Liriope spicatavar produced by a liriope spicatavar GAP base which is a liriope spicatavar origin in Oumiao Town, Xiangyang City of Hubei Province is utilized, and liriope spicatavar steroid saponin is generated through liquid fermentation of liriope spicatavar endogenous fungus strain. The endogenous fungus is an important microorganism for searching new resource of liriope spicatavar steroid saponin.