94 results about "Gene orders" patented technology

The invention discloses gene orders of schistosoma japonicum recombinant multi-epitope antigens BSjGCP-BSj23 and BSjGCP-BSj23-BSj28, a method for expressing and purifying the same, and application thereof in preparing schistosomiasis japonica immunity prevention vaccines and diagnostic reagents. Recombinant multi-epitope nucleic acid vaccines pCMV-BSjGCP-BSj23 and pCMV-BSjGCP-BSj23-BSj28 obtain 14.76 percent and 64.95 percent of worm reduction rates respectively in Kunming mice. The recombinant multi-epitope antigens pGEX-BSjGCP-BSj23 and pGEX-BSjGCP-BSj23-BSj28 obtain 15.7 percent and 57.99 percent of worm reduction rates in immunizing BalB / c mice, and obtain 91.0 percent and 89.9 percent of sensitivities as well as 97.8 percent and 93.4 percent of specificities respectively as diagnostic antigens.

The present invention relates to a gene sequence of an artificially synthesized protein expressing high toxicity to Lepidoptera pests and an application thereof in the field of plant protection, which belongs to the technical field of biological control. The gene sequence of the artificially synthesized protein expressing high toxicity to Lepidoptera pests is characterized in that a coding area is provided with a nucleotide sequence which is 81 percent similar to crylAh gene and can encode the same amino acid sequence as the crylAh gene does. The present invention adjusts the nucleotide composition of the crylAh gene, so that the crylAh gene can approach the codon usage frequency of plant genes but does not change the encoded amino acid sequence; by building the gene sequence obtained by the present invention into an appropriate backbone vector and converting a receptor plant, the gene sequence can be expressed in the plant; the result of a test shows that because of expressing the gene, a positive plant acquires resistibility to Lepidoptera pests.

The invention relates to research field of plant stress resistance gene. The invention provides a peanut aquaporin gene AhAQ1 gene order and a coded protein order thereof. The invention also providesa method for cloning the aquaporin gene AhAQ1 in a peanut. The expression analysis of the aquaporin gene AhAQ1 shows that the expression of the gene is obviously increased in different tissues of thepeanut under the condition of salt stress and also shows that the gene is related to the plant salt resistance. The transgene test analysis also shows that the overexpression of the gene can enhance the salt resistance of the peanut to a certain extent.

The invention discloses an amplimer of a large yellow croaker mitochondrion complete genome sequence and an application thereof. The 15 pairs of amplimer are applied, a gene order is respectively shown as SEQ ID No: 1-30, the amplimer is further used for obtaining a Tai Qu family large yellow croaker mitochondrion complete genome sequence and a Min-Yue East family large yellow croaker mitochondrion complete genome sequence which are respectively shown as SEQ ID No: 31 and SEQ ID No: 32, and an amplimer of large yellow croakers in a high variation region is obtained. The amplimer and the application have the advantages that the Tai Qu family large yellow croaker and Min-Yue East family large yellow croaker mitochondrion complete genome sequences can be quickly and efficiently obtained respectively through the 15 pairs of amplimers; and the amplimer of the large yellow croakers in the high variation region provides support for the research of species identification, geographical population identification, germ plasm resources and genetic diversity of the large yellow croakers, and further provide the basis for developing an identification technology of Tai Qu family large yellow croakers and Min-Yue East family large yellow croakers.

The invention relates to gene order and amino acid component of (S)-carbonyl reductase of Candida parapsilosis CCTCC M203011, and the (S)-special carbonyl reductase gene can be used in separating recemic compound with biological method, which belongs to biological engineering sphere. The invention is to confirm (S)-special carbonyl reductase of Candida parapsilosis CCTCC M203011, its gene nucleotide sequence is SEQ ID NO:1, and its amino acid component is SEQ ID NO:2. By this gene, the recombination strain can be used in racemic chirality resolution and can arrange conditions for the heterogeneity, conversion approach and reaction mechanism of the microbial stereoselectivity oxidoreductase used in chirality resolution.

The present invention relates to single tube in-situ nested PCR method and its application in nucleotide detection. The reaction liquid contains both inside primer and outer primer, and the inner primer has mismatching with target gene sequence or one section of non-specific gene sequence in the 5' end. The inner primer and the original template have highest annealing temperature lower than that of the first round reaction, while the inner primer and the matched template have the melting temperature higher than the highest allowed annealing temperature of the outer primer. The first round PCR cycle annealing temperature is controlled for only the outer primer to be capable of annealing amplification, and the second round PCR cycle annealing temperature is raised for only the inner primer to be capable of annealing amplification. Between the first round and the second round, there are two or more transition stages with lower circular annealing temperature for the inner primer and the original template to form matching template to start the second round reaction.

The invention relates to a bicistronic mRNA coexpression gene transporter and a preparation method thereof. The gene order of the gene transporter is SEQ ID No. 15; the gene transporter from 5' end to 3' end comprises bovine Nuclear matrix binding region MAR, artificially constructed combined promoter CAG, Profilin gene FSTN, internal ribosome entry site (IRES), green fluorescent protein AcGFP gene and Rabbit globin poly A signal region; the preparation method comprises the following steps: constructing a carrier pCAG-IRES2-AcGFP1; obtaining FSTN gene and inserting the pCAG-IRES2-AcGFP1 carrier; obtaining the sequence of MAR and inserting the pCAG-IRES2-AcGFP1 carrier; constructing a plasmid carriers so as to obtain the bicistronic mRNA coexpression gene transporter. The gene transporter is not only pure and safe gene transporter but also realizes dual-gene coexpression in any combination, achieves the purpose of multi-gene coexpression through repeated utilization of IRES, and provides new thinking and route for improving the shape of dual or multiple gene control such as economic character.

The invention relates to a hepatitis B virus nucleic acid vaccine optimized by codon. In the invention, hepatitis B surface antigen (HBs) gene order (adr subtype) is analyzed to find codon locus which tells the differences between the codon preferences of the gene order and the codon preferences of the mammal; the codon of the HBs gene order is replaced to obtain the surface antigen gene; the gene order is combined and expanded to obtain MHBs, Pst I, BamH I, double digestion MHBs gene and carrier pSW3891 plasmid optimized by the codon; 10ul connection system is configured to obtain a middle protein gene. The nucleic acid vaccine of the invention overcomes the defects that the differences between prokaryote and eukaryote in terms of codon preferences cause that the foreign gene can not be expressed effectively in mammal reservoir and can not generate relatively good immune sheltering effect; in addition, the invention remarkably improve protein expression of the foreign gene in the mammal reservoir, effectively stimulates immune system of the reservoir to generate relatively good immunological reaction of human body fluids and cellular immune response.

The invention relates to a chloroplast simultaneity expression seven exogenous genes vector forming method. It belongs to genetic engineering technique field. It includes the following steps: forming a chloroplast expression vector containing faeC, faeD, faeE, faeF, faeG, faeH, and faeI seven gene orders which form K88ac flagellin gene cluster; extracting coding K88ac wild bacterium C83907 plasmid; using primer 1 whose nucleotide sequence is same with SEQ ID NO: 1 and primer 2 whose same with SEQ ID NO: 2 to process PCR amplification; nucleotide sequence 3018bp is same with SEQ ID NO.5; using primer 3 whose same with SEQ ID NO:3 and primer 4 whose same with SEQ ID NO:4 to process PCR amplification; the nucleotide sequence is same with SEQ ID NO.6; connecting with pMD18-T vector; transforming to escherichia coli DH5 alpha; and gaining escherichia coli clone pTEI with faeCD sequence. The invention overcomes the key defect of polygene technique of plant genetic improvement, and offers a good method of producing effective antibody plant bacterin for baby pig.

The invention relates to the gene mutation relevant to the paralysis agitans and the detecting method, comprising: extracting DNA sample; designing primer; expanding with PCR; analyzing nucleic acid by means of dHPLC technology; testing the gene order of the sample which is dHPLC positive reaction to definite the genetype; carrying out beta-actin comparison for the sample of repeated extanding negative reaction to eliminate extron deletion mutation. The parkin gene mutation is in the codon promoter of the parkin gene, the second basic group in the exon1, ATG mutated to ACG; on the intersection point of thefifth introne and sixth extron in the exon6, AG mutated to AC; in the sixth extron, the 727th basic group, G mutated to A; in the exon9, the 1016th basic group, C mutated to T; in the position of ring finger intermediate region, in the Znf C6HC zinc region, GCG mutated to CGTC. By applying the gene mutation and the detection method, the detection for the mutation site of the parkin fene is high-effective and low-cost, and the diagnose to the potential PD patient can be made in an early stage.

The invention relates to a heuristic breadth-first searching method for cancer-related genes. According to the method, appearance frequencies of genes in a selected gene subset are used for measuring the genes, and genes with higher appearance frequency are considered as the most important cancer-related genes, on the basis, a classifier is designed and a gene ordering method based on HBSA is established. As proved by study, information gene selection plays an important role in improving the classification performance, and the genes can be probably taken as important tumor clinical diagnosis signs, so discovery of the minimum gene subset with the highest classification performance is a very important research objective. As indicated by experimental results, the heuristic breadth-first searching method can not only obtain favorable generalization performance but also discover important tumor genes. And the relationship of the appearance frequencies of the selected genes and the gene number conforms to power-law distribution. The genes in the gene subset with extremely high classification accuracy are in close relationship with specific tumor subtypes, and even the genes are important genes directly related with the tumor.

The invention provides a recombined endo-chitinase gene sequence based on Pichia pastoris codon preference and a recombined vector containing the recombined gene. The invention adopts modern biotechnology and makes use of PCT so as to synthesize the recombined endo-chitinase gene sequence based on Pichia pastoris codon preference, and then clones the gene sequence in a vector pPIC9K so as to obtain a eukaryon expression vector pSECH. The transformation of Pichia pastoris GS115 is carried out through adopting an electric shock method, and quick screening of positive cloning transformant with higher resistance is carried out according to growth conditions, thereby realizing efficient industrialized production of endo-chitinase at low cost. Moreover, the recombined endo-chitinase gene sequence and the recombined vector have wide application prospect.

The invention discloses an anti-SARS coronavirus cell vaccine and its application, said cell vaccine (1) containing all or part gene fragment of SARS coronavirus S protein all length gene order and 5 specific eukaryotic expression vector of cDNA fragment; (2) expressing the heterogenetic antigen presenting cells of said SARS coronavirus gene corresponding protei; (3) inactivating the heterogenetic antigen presenting cells expressing SARS coronavirus related protein. Use said cell vaccine for immuning host containing humans and rodents such as mice in order to making them generate protective humoral immunity and cellular immunity and resisting the SARS coronavirus infection which causes atypical pneumonia contagion. The security and stability of the invention are fine, production is convenient and price is low, it can induce the body generating the effective humoral immunity and cellular immunity of SARS coronavirus causing atypical pneumonia at the same time; provide effective and cheap vaccines for treating and preventing SARS.

The invention discloses a kind of primer and fluorescent probe that are used for sulfate-reducing bacteria detection, relating to primer and fluorescent probe that are used for microbe detection. It overcomes shortcomings of long detection time, high non-specificity and smaller detection result compared to correct result which are existed in current quantitative determination method for SBR bacteria. The gene order of upstream primer is 5'-GTTCCCTGCTCGTGCCCT-3'and gene order of downstream primer is 5'-TTCCTTGAAGAAGATGTACGGGTT-3'. The fluorescent probe comprises report fluorophor, quenching fluorophor and probe gene order; the probe gene order is 5'-AATGGTGGATGGAAGAAGGC-3', the report fluorophor marked by 5'end in probe gene order is FAM, and the quenching fluoropho marked by 3' end in probe gene order is TAMRA. The invention is characterized by high specificity of primer and fluorescent probe, short SBR bacteria determination time, which needs only 6 hours for sample mould DNA extraction to fluorescence quantitative PCR detection.

Sputum is a main sample for clinical test of tuberculosis, but the prior sputum processing method kills mycobacteria to different degrees during the processing process, thereby detectable rate of the tuberculosis mycobacteria is severely reduced. The invention firstly provides a method for quick and effective liquefaction of the sputum, has the characteristics of simple operational procedures and good liquefaction effect, and has the advantage that the method has no killing function on the mycobacteria in the sputum during the liquefaction process. The invention also provides a test method for tuberculosis mycobacterium complex nucleic acids with high sensitivity and strong specificity, wherein, specific amplified products of Ag85B gene are tested through fluorescent PCR amplification method and specific primers and a Taqman fluorescent probe are designed according to a tuberculosis mycobacterium complex Ag85B gene order, thereby the tuberculosis mycobacterium complex is tested. The invention is implemented by the two mutually independent and mutually associated inventions, and provides a method for testing the tuberculosis mycobacterium complex in clinical samples, which has the advantages of high efficiency, quickness, high sensitivity and strong specificity.

The invention discloses a C2H2 type zine finger protein gene AhZFP1 gene order in peanut, responding to salt stress, and coded protein order thereof. The invention also provides a method for cloning the C2H2 type zine finger protein gene AhZFP1 gene. The expression analysis of the C2H2 type zine finger protein gene AhZFP1 shows that the expression of the gene is obviously increased in different tissues of the peanut under the condition of salt stress, and a transgene test analysis also shows that the overexpression of the gene can surely enhance the salt resistance of the peanut.

The invention relates to NuBCP-9 and Tumstatin(74-98) fusion genetic engineering antitumor peptide which is designed to be synthetic primer according to colon bacillus favorable codon, NuBCP-9 and Tumstation(74-98) fusion peptide gene order connected by flexible peptide (G4S3) is obtained by PCR through an SOE method, the NuBCP-9 and Tumstation(74-98) fusion peptide forms recombination expression plasmid after connecting with a carrier Pet 32A(+), the recombination expression plasmid is transferred to colon bacillus BL21 for performing the soluble efficient expression and purifying and enzyme-cutting to obtain the purpose peptide. The fusion peptide has better inhibition function to skin cell proliferation in umbilical vein and lung cancer cell, primarily shows the multitarget antitumor effect, and can have better clinical application prospect.

The invention discloses a gene order fragment fast positioning method based on a bitmap; the method comprises the following steps: constructing the bitmap, circulating and excursing each gene order fragment in a gene reference chain one by one so as to form a plurality of gene order fragment vectors, mapping the vectors to the only data bit in the bitmap, replacing 0 by 1, counting 1 appearing frequency t before the column on which the line located, and storing a key value in a t position of a Hash bucket corresponding to a line number R of the data bit in the database. If all data bits are set by 1, adding the data bits into a clash assembly; circulating and excursing to-be-positioned gene order fragments one by one so as to form a plurality of gene order fragment vectors, mapping the vectors to the only data bit in the bitmap, thus positioning a Hash data table. The positioning method can fast filter untargeted query data, can provide relatively accurate data positions of the Hash bucket, thus greatly accelerating a Hash bucket dividing mode organizing data query process; the gene order fragment fast positioning method based on the bitmap is high in space efficiency, fast in positioning speed, and accurate in positioning.

The invention relates to a very important protein gene PTP2 related to propagation and infection of nosema bombycis nageli. A gene order CDS for encoding nosema bombycis nageli polar tube PTP2 is obtained according to 9-fold genome data of the nosema bombycis nageli and proven by a series of biotechnology clonings. The nosema bombycis nageli polar tube PTP2 obtained in the invention plays a very important role in studying a propagation and infection mechanism of microsporidian, thus further playing a role in productive practice and prevention and treatment of the microsporidian disease. Experiments and analyses such as RNAi intervention and the like indicate that after the polar tube protein PTP2 is intervened, the microsporidian loses the capacity of infecting a host, therefore, the polar tube protein PTP2 of the microsporidian and potential regulator genes thereof become new factors for treating microsporidian infection, in addition, the method of modifying genes can be adopted to express the polar tube protein PTP2 in an abundant way, thus killing part of pests such as locust and the like.

The invention relates to a protein, which is characterized by having the function of resisting insect blood cell immunity based on cloning peptide sequence of an epidermal protein ENO1p of format nematodes (Steinernema glasri) to an epidermal protein gene enol. The invention also relates to a method for preparing the protein and polynucleotide thereof and application thereof.

The invention provides a recombination lactic acid galactococcus method of expessing pig transmissible gastroenteristis virus mutation S protein, which comprises the following steps: designing double primers according to the total gene order of pig Transmissible gastroenteristis virus S protein and gene fusion characteristic of expressing the carrier plasmid; proceeding PCR; getting the 2007bp destination fragments of four main antigen sites with TGEV S gene; connecting with the carrier plasmid PNZ8112 of secretory expression; getting into the host lactic acid galactococcus NZ9000 cell by electric conversion; expressing under the inducing of nisin. The invention constructs the expression carrier system of coronaviridae TGEV S mutein lactic acid galactococcus, which expresses about 66KDa destination protein of four main antigen sites with TGEV. Immunological ink mist experiment and indirect immunological fluorescein test indicate that the exogenesis protein is able to react with TGEV immunological serum and the recombination S protein has the same antigenic as TGEV natural antigenic.

The invention relates to the method used PCR to identify the goat early embryo sex. It includes the following steps: sampling from goat early embryo sex to make template DNA; designing primer according to goat SRY gene order; adding the template DNA, primer, DNA polymerase, reaction buffer solution, and dNTP into reaction tube to use as reactant; repeating denaturalization-annealing-stretching three steps to the end of the reaction. Its features are that it adds ethidium bromide into the reaction tube before PCR, directly observes the result after PCR by putting the reaction tube under ultraviolet lamp. It designs a pair of primers according to goat Y chromosome specificity sequence, identifies by non-electrophoresis PCR, simplifies operational programs, and is convenient for popularization and application in production.

The invention discloses a method for using silkworm cultured cell to express antibacterial peptide Cecropin B. The method comprises the following steps: (1) using the total RNA extracted from the fat body cells of wild silkworm chrysalis as template, adopting RT-PCR amplification to obtain wild silkworm antibacterial peptide Cecropin B gene; using 1% agarose gel electrophoresis to perform PCR product analysis to the antibacterial peptide Cecropin B gene, using a PCR purification kit of Qiagene for purification, then cloning the purified PCR product to TA vector pCR2.1 to obtain pCR2.1-Cecropin B, utilizing the dideoxynucleotide chain termination to confirm the correctness of cloned gene order; using restriction endonuclease BamHI and HindIII to digest pCR2.1-Cecropin B and obtain Cecropin B genetic fragments, then cloning rhabdovirus transfer vector pBlueBacHisa in the genetic fragments to obtain reconstituted transfer vector; performing cotransfection of the reconstituted transfer vector and silkworm wild-type nuclear polyhedrosis virus BmNPV DNA in silkworm cultured cell, performing homologous recombination to generate recombinant virus; and (3) inoculating the recombinant virus containing wild silkworm antibacterial peptide Cecropin B gene in the silkworm cultured cell, infecting at 27 DEG C for three days to express the infection, and centrifuging to collect silkworm cultured cell.

The invention discloses a kind of primer and fluorescent probe that are used for anaerobic denitrifying bacteria detection, relating to primer and fluorescent probe that are used for microbe detection. It overcomes shortcomings of long detection time, smaller detection result compared to correct result, complicate operation and high detection price existed in current quantitative determination method. The gene order of upstream primer is 5'-ATGCGTAGCCGACCTGAGA-3'and gene order of downstream primer is 5'-CGTCAGACTTTCGTCCATTGC-3'. The gene order of fluorescent probe is 5'-CGGGAGGCAGCAGT-3'; the report fluorophor marked by 5'end in fluorescent probe is FAM, and the quenching fluoropho marked by 3' end is TAMR- MGB. The invention provides specific primer and probe for anaerobic denitrifying bacteria detection, and it is characterized by accurate bacteria counting, simplified operation, reduced cost, shortened detection period and practical meaning in production.

The invention relates to a human KGF- 2 mutant, the amino sequence is demonstrated as sequence 2 in sequence table and the gene order is the squence 1 in squence table. The invention aslo provides the expression vector and engineering bacteria containing mutant gene. The invention aslo provides the method for preparing said KGF-2 mutant, comprising: (1) culturing bacillus coli engineering bacteria at proper condition, and inserting gene order in multiple coloning point of expression vector to express human KGF-2 mutant; (2) separating and purifying the human keratinized cell growing factor -2 mutant through column chromatography.

The invention discloses a molecular tagged method of indicating and identifying the size of a qin-chuan cattle body by using GDF5 gene, which comprises the following steps: designing a pair of primers according to the cattle GDF5 gene order; utilizing the primers to perform PCR amplification to the genom DNA of the cattle; utilizing the PCR product gained and digested by restriction enzyme Mva I; and then using 2.5 percent of sepharose gel to perform the electrophoretic separation to the digested PCR product, detecting T / C variable sites of a 586 site of GDF5 gene exon 1; and respectively naming the variable sites of the GDF5 gene exon 1 of the cattle as C base, T base and T / C hybrid base. According to the experiment result, the length of the cattle with TT gene is 5.64-6.87cm less than that of the cattle with CC gene; and the hip width of the cattle with TT gene is 3.46-4.00cm less than that of the cattle with CC gene. The method has simple operation, low cost, high accuracy and rapid detection.

The invention discloses a group of gene sequence of gastric cancer auxiliary diagnosis and a gene chip and a kit aiming at the group of gene sequence. The invention also discloses the application of the group of gene sequence of gastric cancer auxiliary diagnosis. The gene sequence of gastric cancer auxiliary diagnosis applied in the invention can type gastric cancer from molecular level and helps clinical diagnosis, thus more effectively healing patients and reducing treatment side effect to the utmost extent.

The invention relates to a cDNA sequence, a coding protein, a full-length gene sequence and the clone method of a 26S proteinase body subunit RPT2 gene (DvRPT2)in Dunaliella viridis. The uplift expression of 26S proteinase body subunit RPT2 gene in Dunaliella viridis enhances the hydrolytic efficient of TP, accelerates that the substrate protein enters the active position of the proteinase body, reduces the abnormal protein content in the cell, and improves the salt resistance of Dunaliella viridis, so the research about Dunaliella viridis RPT2 subunit indicates the protein degradation pathway of 26S proteinase body, the salt endurance mechanism and the salt-resistance plant improvement, which has the finite theoretical and actual application value.

The invention relates to a very important protein gene PTP3 related to propagation and infection of nosema bombycis nageli. A gene order CDS for encoding nosema bombycis nageli polar tube PTP3 is obtained according to 9-fold genome data of the nosema bombycis nageli and proven by a series of biotechnology clonings. The polar tube PTP3 obtained in the invention plays a very important role in studying a propagation and infection mechanism of microsporidian, thus playing a role in productive practice and prevention and treatment of the microsporidian disease. Experiments and analyses such as RNAi intervention and the like indicate that after the polar tube protein PTP3 is intervened, the microsporidian loses the capacity of infecting a host, therefore, the polar tube protein PTP3 of the microsporidian and potential regulator genes thereof become new factors for treating microsporidian infection, in addition, the method of modifying genes can be adopted to express the polar tube protein PTP3 in an abundant way, thus killing part of pests such as locust and the like.