168 results about "Transcription (biology)" patented technology

The invention discloses a detection kit and detection method for H1, H3 and H9 type avian influenza viruses and belongs to the technical field of biology. The kit comprises an RNA extraction kit, an RNA reverse transcription kit and an RPA detection kit. The RNA extraction kit is used for extracting RNA of a to-be-detected sample to be used as a detection template. The RNA reverse transcription kit is used for conducting reverse transcription on the RNA in the detection template to obtain cDNA. The RPA detection kit comprises primers and fluorescent dye. The primers are used for amplifying the cDNA to obtain an amplified DNA sequence. The primers comprise the forward primer and the reverse primer. The fluorescent dye is used for conducting fluorescence detection on the amplified DNA sequence. According to the detection kit and the detection method, the avian influenza viruses can be rapidly, easily and conveniently detected.

The invention relates to a method for realizing efficient and fixed-point transgenosis by using a TALEN (Transcription Activator-Like Effector Nuclease) technology, belonging to the field of molecular biology. The method comprises the following steps of: designing a TALEN target point with a specific sequence in a special position in an organism genome by using the TALEN technology, constructing a TALEN plasmid, and meanwhile, constructing an exogenous gene plasmid; then, co-transfecting the TALEN plasmid and an exogenous gene into recipient cells, and then incising in a to-be-integrated position of a recipient cell genome; and finally, automatically inserting an exogenous gene sequence to the exogenous gene at the incision of the recipient cell genome. The fixed-point integration of the exogenous gene on the genome is realized by using the method; and the method is simple in operation, high in controllability and strong in practicability.

The invention relates to the technical field of molecular biology, and particularly discloses a sarcoma fusion gene joint detection primer group, kit and detection method. The kit provided by the invention comprises: 1) a special linker for DNA library construction; 2) a specific composite primer for detecting fusion genes and point mutation; and 3) a composite primer for library amplification. DNA and RNA of a to-be-detected sample are respectively extracted, reverse transcription is performed on RNA to synthesize cDNA, the cDNA is mixed with DNA, fragmentation, terminal repair and linker ligation are performed on the mixture, PCR amplification is performed on the product, a library is enriched, and high-throughput sequencing is performed. According to the fusion gene detection kit and detection method provided by the invention, various fusion genes or mutations related to sarcoma can be detected at one time, the detection efficiency is improved, the operation is convenient, the consumption time is short, and the sensitivity is relatively high and can reach 0.1%.

The invention belongs to the technical field of crop disease diagnosis and molecular biology and discloses a multiple RT-PCR (Reverse Transcription-Polymerase Chain Reaction) method for synchronously detecting five watermelon viruses. A PCR system contains five pairs of specific primers (SEQ ID NO1-10), and the five viruses are synchronously multiplied in one PCR system, so as to obtain specific bands 1464bp, 1023bp, 810bp, 600bp and 327bp and establish and optimize a multiple RT-PCR reaction system capable of synchronously detecting the viruses CiLCV, CGMMV, ZYMV, MABYV and WMV. A detection result indicates that the optimized multiple RT-PCR reaction system can detect a field sample quickly, efficiently and accurately. The method has a significant meaning for forcasting of occurrence and disease epidemiology of five common viruses such as CiLCV and the like on monitored watermelons.

The invention discloses preparation, detection and application of a polyclonal antibody of a Yam mild mosaic virus (YMMV). Based on a YMMV genome, the inventor purifies a nuclear inclusion protein/coat protein (NIb/CP) of the YMMV through prokaryotic expression and uses the nuclear inclusion protein/coat protein to prepare the polyclonal antibody for an immune rabbit. The polyclonal antibody can specifically identify the Nib/CP protein of YMMV through prokaryotic expression and the Nib/CP protein of the YMMV virus which infects common yam rhizome, and generates specific immunity response. On the basis, the inventor further builds a set of efficient, high sensitive and accurate IC-RT-PCR (Immunocapture-Reverse Transcription-Polymerase Chain Reaction, ELISA (Enzyme-Linked Immuno Sorbent Assay) and Western blot methods which can specifically detect the YMMV from a common yam rhizome plant which is infected by YMMV. By applying the invention, a material basis and a technical support can be provided for research on interaction of the YMMV and the host, quick detection of the virus and somatotype and molecular biology study, thereby laying a solid foundation for epidemiologically monitoring and preventing treating the virus.

The invention relates to a plant expression vector and a construction method thereof, and a method for producing chicken alpha interferon by utilizing crowtoe as a bioreactor, belonging to the technical field of biology. The plane expression vector pSFRLH is shown as the figure 1 in the specification. In the invention, main stem sections, rachis stem sections and tender leaves of crowtoe are used as explants, and a constructed plane expression vector chicken alpha interferon gene ChIFN-alpha is subjected to genetic transformation and regeneration and then subjected to PCR (Polymerase Chain Reaction), RT-PCR (Reverse Transcription-Polymerase Chain Reaction), Southern blot, Western blot and ELISA (Enzyme-Linked Immuno Sorbent Assay). Proved by a detection result, the ChIFN-alpha gene is integrated into a crowtoe chromosome genome and correctly transcribed, translated and expressed, and recombinant protein can be neutralized by a ChIFN-alpha antibody. The invention lays the foundation for producing animal medical protein by utilizing crowtoe pasture grass as a bioreactor and provides a new thinking for the control of animal epidemic diseases.

The invention provides rationally designed multi-targeting therapeutic agents for myotonic dystrophy type 1 (DM1), an incurable neuromuscular disease that originates in an abnormal expansion of CTG repeats (CTG^exp) in the DMPK gene. The rationally designed small molecules target the DM1 pathobiology in three distinct ways: (1) binding the expanded trinucleotide repeat, CTG^exp, and inhibiting its transcription to the toxic CUG^exp RNA, (2) binding the CUG^exp RNA and releasing sequestered muscleblind-like protein (MBNL1), and (3) cleaving the toxic CUG^exp in an RNase-like manner. Importantly, the compounds can reduce the levels of CUG^exp in DM1 model cells and reverse two separate CUG^exp-induced phenotypes of DM1.

The invention relates to a complementary DNA (cDNA) sequence of tobacco NtFT1 genes and transient expression thereof for inducing tobacco early blossoming, in particular to NtFT1 genes of tobacco FT (Flowering locus T) and application of the NtFT1 genes in inducing the tobacco early blossoming, and belongs to the field of genes in molecular biology. The invention discloses a full-length coded sequence of the NtFT1 genes derived from tobacco for controlling the plant blossom time and a protein sequence coded by using the full-length coded sequence. The NtFT1 genes are obtained by performing Blastn comparison and splice site analysis on the data searched from China tobacco genome sequencing planning data through homologous sequence searching. According to the cDNA sequence, a method for performing semi-quantitative polymerase chain reaction (PCR) or reverse transcription-polymerase chain reaction (RT-PCR) on NtFT1 gene specific primers is designed to clone the genes. According to the invention, a potato virus X (PVX) virus expression vector of the NtFT1 genes is also constructed, and the constructed virus expression vector is used for performing diafiltration and infestation on flue-cured tobacco varieties including Hongda and Yunnan tobaccos 87 and K326 through agrobacterium, and can induce the early blossoming of the varieties to prove that the genes have a function of inducing the early blossoming of cured tobaccos. When a PVX virus transient expression system of the NtFT1 genes is used for tobacco breeding, the growing time of the tobaccos can be shortened, and the fixed number of years for breeding a new tobacco variety is reduced.

The invention discloses a method for testing the development process of silkworm embryos. The method specifically comprises the following steps of: (1), establishing a model expressing the incidence relation between the relative transcription expression quantity of the acetylcholine esterase type 1 (ace1) of a to-be-tested variety of the silkworm embryos and the development process of the embryos; (2), selecting at least one embryo from the to-be-tested silkworm embryos to use as a sample, testing the relative transcription expression quantity of the ace1 of the sample, and then contrasting with the model obtained in step (1) to determine the development process of the silkworm embryos according to the relative transcription expression quantity of the ace1 of the to-be-tested sample. The development stage of a whole graine can be accurately determined by the method through the test on the development process of a small quantity of silkworm embryos. Manual operation errors can be reduced, the test process is simplified, and a development process identification method is provided for molecular biology study of the silkworm embryos.

The invention discloses a one-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection kit for an influenza virus, relating to a PCR kit in the technical field of biology. The kit comprises one-step 5xRT-PCR Buffer, positive control, negative control, three pairs of specific primers and Enzyme Mix; a specific primer AF: 5'-ATTCTAACCGAGGTCGAAACGT-3' and a specific primer AR: 5'-GACAAAGCGTCTACGCTGCAGTC-3' are designed according to a gene M of an influenza virus type A; a specific primer is designed according to a gene NS of an influenza virus type B; and a specific primer is designed according to a gene HA of an influenza virus A type H1N1. The detection kit has a short detection time period, high detection efficiency, high virus detection specificity, high accuracy, higher detection sensitivity than an immunological detection method and high experiment result repeatability, can be applied to qualitative analysis of viruses, and is easy to operate and popularize.

The invention discloses a multiple RT-PCR (reverse transcription-polymerase chain reaction) detection primer group and kit for rapidly distinguishing PEDV (porcine epidemic diarrhea virus), PDCoV (porcine deltacoronavirus) and PReoV (porcine reovirus), and belongs to the technical field of animal virology and molecular biology. The kit comprises primer sequences as shown in SEQ ID 1-6, Prime STARHR buffer solution, a cDNA template and sterile water. The multiple RT-PCR detection kit of the PEDV, the PDCoV and the PReoV is amplified by multiple RT-PCR, three lesions are similar and serve as primary detection of RNA viruses, total detection time is controlled to be about 2 hours, the detection method is easy to operate, convenient and rapid, and rapid detection can be achieved.

The invention relates to the field of molecular biology, and relates to a circular RNA detection method and kit. The circular RNA detection method comprises the following steps: S1, primer design; S2,reverse transcription reaction: taking 0.1-1 [mu]g of extracted total RNA, adding N6 Random Primer and RNase-Free H2O, performing mixing to obtain a reaction solution I, adding 5 x HT RT Buffer intothe reaction solution I, adding HT M-MLV Mix into the reaction solution I, and finally adding RNase-Free H2O to obtain cDNA; and S3, HS PCR amplification: performing qPCR detection by using the FSjodPrimers obtained in the step S1, 2 x SNP Taq Mix, 50 x ROX Mix *, the RNase-Free H2O and the cDNA obtained in the step S2.

The invention discloses a double RT-PCR (reverse transcription-polymerase chain reaction) detection method of CVB (chrysanthemum virus B) and CChMVd (chrysanthemum chlorotic mottle viroid), belonging to the virus detection field of molecular biology. The method comprises the following steps of: (1) sampling from chrysanthemum plants, and extracting total RNA (ribonucleic acid); (2) respectively designing two pairs of specific primers for CVB and CChMVd: CVB-F and CVB-R; CChMVd-F and CChMVd-R; (3) carrying out reverse transcription by taking a random hexamer as a primer to obtain two kinds of cDNA (complementary deoxyribonucleic acid); and (4) amplifying the cDNA by utilizing the RT-PCR technology, and carrying out agarose gel electrophoresis on the amplified products to obtain a 665bp specific fragment for expressing infection of CVB and obtain a 206bp specific fragment for expressing infection of CChMVd. The detection method provided by the invention can be used for simultaneously detecting diseased plants which are compositely infected by CVB and CChMVd, and has the advantages of strong detection specificity, high sensitivity, simple process and cost saving.

The invention discloses a primer combination and plasmid for simultaneously detecting 9 kinds of respiratory tract viruses and a detection kit and belongs to the technical field of biology. The primercombination comprises primer pairs for detecting an influenza a virus, an influenza b virus, an H7 influenza virus, an NL63 corona virus, a 229E corona virus, a type 2 parainfluenza virus, Bocavirus,a respiratory syncytial virus A, a respiratory syncytial virus B and a human internal-reference GAPDH gene. The detection kit comprises a 2*multiple reverse transcription PCR reaction solution, a reverse transcription PCR enzyme mixed solution, a primer mixed solution, non-RNAase water, a magnetic bead mixed solution, a reported buffer solution, a positive control and phycoerythrin-modified streptavidin. According to the primer combination and plasmid and the detection kit, by virtue of a liquid-phase chip technology, the manufactured detection kit can be used for carrying out qualitative analysis on a variety of respiratory tract viruses simultaneously, and the sensitivity and specificity of detection reach PCR levels; and types of respiratory tract virus infections can be rapidly and accurately judged, early etiologic diagnosis and treatment on clinical respiratory tract infections are facilitated, the shortening of a treatment cycle is facilitated, the medical expense is reduced, and good social benefits are generated.

The invention relates to the technical field of biology and discloses a whole genome cloning method of pelteobagrus fulvidraco bacillus-shaped virus. The method comprises the following steps of: designing 14 specific primers for reverse transcription and 14 pairs of specific primers for PCR, wherein the primer sequences are shown as SEQ ID No.1-SEQ ID No.14; extracting pelteobagrus fulvidraco bacillus-shaped virus RNA; taking the extracted viral RNA as a template, respectively adding the extracted viral RNA into a reverse transcription reaction system and a PCR reaction system by using the primers, cloning the whole genome of the pelteobagrus fulvidraco bacillus-shaped virus by sections under reaction conditions, wherein 13 fragments of the amplified product are 2000bp in size, one fragment is 1000bp in size, a total of 14 fragments contain the entire genome sequence of the pelteobagrus fulvidraco bacillus-shaped virus. The invention provides a cloning method of the whole genome of thepelteobagrus fulvidraco bacillus-shaped virus. The method not only can be used for rapid cloning of the whole genome of the pelteobagrus fulvidraco bacillus-shaped virus, but also can be used for researching molecular biology pathogenic mechanism and molecular epidemiology investigation of the pelteobagrus fulvidraco bacillus-shaped virus, and is convenient to use and has good effect.