68 results about "Gene Component" patented technology

The invention belongs to the field of cells and in particular relates to an efficient induced pluripotent stem cell reprogramming method for blood cells. The method comprises the following steps: S1,extracting monocyte from a blood specimen, and performing selective culture with an amplification culture medium so as to obtain erythrocyte progenitor cells; S2, introducing a free carrier with at least one potentiality determinant factor into the obtained erythrocyte progenitor cells; S3, culturing the erythrocyte progenitor cells with the free carrier by using a pluripotent stem cell inductionculture medium, and performing induction in a feeding layer free system so as to obtain reprogrammed intermediate state cells; S4, after induction is completed, replacing the pluripotent stem cell induction culture medium in the step S3 by a pluripotent stem cell culture medium to maintain culture, thereby obtaining cells that potentiality determinant factor expression is vanished and expression of endogenous pluripotent genes POU5F1, NANOG, TRA-1-60 and TRA-1-81 is activated, namely the induced pluripotent stem cells. The method has the beneficial effects that pluripotent stem cells without endogenous gene components can be efficiently induced, and the method is applicable to preclinical study and clinical application.

The invention provides a computer implemented process for constructing a scalable output network model of a bioparticle. The process includes computer implemented steps of: (a) accessing a database of network gene components including an annotated network set of open reading frames (ORFs) of a bioparticle genome; (b) forming a data structure associating the network gene components with network reaction components, the data structure establishing a data set specifying a network model of connectivity and flow of the network reaction components, and (c) transforming the data set into a mathematical description of reactant fluxes defining the network model of connectivity and flow, wherein the mathematical description defines a scalable output network model of a bioparticle.

The invention discloses phage-assisted multi-bacterial continuous directed evolution system and method. The system includes: a phage SM which carries an evolution target gene, a plurality of assistant plasmids HP which support different SM proliferations before and after the evolution, and a plasmid IP which induces mutation. In the system, the different HPs are arranged in different host bacteria, so that proliferation and evolution efficiencies of the phage are effectively improved, and meanwhile, mutual interference between different gene components is avoided. The system and method have great practicability and can be applied to directed evolution of various genes.

In various embodiments, the invention provides expression systems for heterologous protein expression in vegetative plant tissues, utilizing plant seed gene components that are adapted to orchestrate high levels of vegetative protein production. The expression systems may include host plant cells having recombinant genomes, and the plant cells may be maintained under protein expressing conditions, for example in tissue culture. The cells may be induced to express an ABD transcription factor, for example by transformation with a vector having a constitutive ABB expression cassette. The recombinant sequences in operative linkage may include an integrated expression promoter responsive to the ABI3 transcription factor, such as an arcelin gene promoter, a vicilin gene promoter and a napin gene promoter. A 5′ untranslated region may include a region of an ABA responsive plant seed gene or an AB 13 responsive plant seed gene. A plant secretion signal peptide coding sequence may be included. An integrated heterologous protein coding region, encoding a recombinant protein, may be provided in an open reading frame with the signal peptide coding sequence. A 3′ untranslated region may be provided having a polyadenylation signal.

The invention discloses an LAMP detecting primer group, a kit and a detecting method for a cry3A gene in transgenic plant. The primer group consists of 5 specific primers with nucleotide sequences as shown in SEQ ID NO:1-5. The detecting kit comprises detecting primer liquor, loop primer liquor, DNA polymerase with strand displacement activity, 10*reaction buffer liquor, dNTPs liquor, positive DNA contrast and negative DNA contrast. The kit further contains a color developing agent. The detecting method adopts the specific primers and the DNA polymerase with the strand displacement activity to carry out amplification onto a sample DNA template at 63 DEG C-65 DEG C, and adopts a method of adding the color developing agent to observe color changes or to directly observe turbidity change of precipitates in a reaction tube, and the like, to judge whether the sample contains the cry3A gene component or not. The LAMP detecting primer group, kit and detecting method disclosed by the invention do not need a special apparatus, have the characteristics of being quick and efficient, simple and convenient to operate, strong in specificity, high in sensitivity, and the like, and are suitable for site detection.

The invention discloses a method for detecting transgenic components of corn by utilizing quadruple real-time fluorescent PCR, and primers and probes used in the method. The method disclosed by the invention comprises the following steps of designing the primers and the probes required by quadruple real-time fluorescent PCR amplification; extracting genome DNA in the corn to be detected; carryingout quadruple real-time fluorescent PCR amplification; and judging whether the corn to be detected is transgenic corn or not according to a PCR amplification result. The invention provides the effective method for rapidly detecting whether the corn and a deep processing product thereof are the transgenic corn or not.

The invention discloses a detection method for soybean transgenic ingredients, and belongs to the technical field of biology. The method includes the steps that the transgenic ingredients, endogenesis standard genes and exogenous standard genes are determined; a multiple amplification primer is prepared; a mixed sample is obtained through sampling and mixing; a genome of the mixed sample is extracted, and the exogenous standard genes are added to obtain mixed nucleic acid; a high-throughput sequencing library is constructed; high-throughput sequencing is conducted on the high-throughput sequencing library to obtain a sequencing fragment set; the number of sequencing fragments of the transgenic ingredients, the number of sequencing fragments of the endogenesis standard genes and the number of sequencing fragments of the exogenous standard genes in the sample to be detected are obtained; according to the number of the sequencing fragments of the exogenous standard genes, whether an experiment is successful or not is judged, and the content of the transgenic ingredients in the sample is calculated; according to the content of the transgenic ingredients, whether the sample to be detected contains content of the transgenic ingredients or not is judged. Any kind of transgenic ingredients in the sample to be detected can be quantitatively detected at the same time.

The invention belongs to the field of cells and particularly relates to a method using salamander Oct4 to reprogram human blood cells into iPSC. The method includes the steps of S1, acquiring progenitor red blood cells; S2, importing a free vector containing a salamander Oct4 transcription factor and other transcription factors into the progenitor red blood cells obtained in S1; S3, subjecting theprogenitor red blood cells, containing the free vector, obtained in S2 to multipotential stem cell induction medium culture, and inducing into reprograming intermediate-state cells in a feed-layer-free system; S4, after complete induction, replacing the multipotential stem cell induction medium in S3 with multipotential stem cell culture medium to continue the culture so as to obtain the cells with disappeared salamander Oct4 transcription factor and other transcription factor expression and activated endogenous multipotential gene POU5F1, NANOG, TRA-1-60 and TRA-1-81 expression, wherein thecells are the iPSc. The method has the advantages that efficient induction can be performed to generate the hiPSC without exogenous gene components, and the method is applicable to preclinical study and clinical application.

The invention discloses a gene targeting vector based on poultry adeno-associated virus, and a construction method thereof. The vector takes the poultry adeno-associated virus as a framework, wherein the inner sides of 5' and 3' LTR are connected with two homologous arms respectively; a gene component with positive selective resistance is connected between the two homologous arms; and two insulator elements are arranged between the gene component with positive selective resistance and the homologous arms respectively. The construction method comprises the following steps: (1) constructing a gene transfer vector of the poultry adeno-associated virus; (2) constructing pMD19-2HS4 plasmid; (3) amplifying OV complete sequence; (4) amplifying every component of the gene targeting vector and performing PCR splicing; and (5) constructing the gene targeting vector. The gene targeting vector has the advantages of effectively inhibiting position effect in a gene targeting process, improving the efficiency of screening positive clones, inhibiting random integration, improving the homologous recombination efficiency of the positive clones and having high gene transfer efficiency for non-active genes or temporal specific expressed genes.

The invention discloses a transgenic ingredient high-throughput screening method, a detection kit and application thereof. The method comprises the following steps: according to the condition of exogenous insertion genes in commercialized transgenic crops, selecting 10 fragments capable of covering all commercialized transgenic strains as screening elements aiming at the exogenous genes, and obtaining conserved sequences of the 10 screening elements through bioinformatics analysis; a DNA primer pair designed aiming at a detection target and a corresponding TaqMan probe are embedded in each micropore of the microfluidic chip in advance, specific primers and probes are designed according to a sequence to carry out microfluidic chip detection, and whether a sample contains transgenic ingredients or not and which crop contains the transgenic ingredients are judged according to an amplification result. The sensitivity test of the method reaches 0.1%, and the method has good specificity. The kit and the detection method provided by the invention provide a good application prospect for high-throughput screening and detection of transgenic ingredients in prepackaged and processed food.

The invention relates to the technical field of biological detection, and provides a NASBA-ELISA rapid detection kit for the anti-glufosinate resistance gene component in transgenic corn, wherein theNASBA-ELISA rapid detection kit comprises specific primers designed according to the mRNA sequence of an anti-glufosinate resistance gene bar, a capture probe, a detection probe, an optimized NASBA amplification system and an optimized NASBA product detection system, can quickly and easily detect the anti-glufosinate resistance gene component in corn, has advantages of high specificity and easy promotion, and can further help the early diagnosis of the anti-glufosinate resistance gene component bar in transgenic corn. The detection method comprises: designing and artificially synthesizing specific primers for the mRNA sequence of an anti-glufosinate resistance gene bar, a capture probe and a detection probe, determining a NASBA amplification reaction procedure and a NASBA product detectionsystem, and other steps. According to the present invention, the detection method can avoid the requirements on special equipment and professionals, and further has advantages of high sensitivity andgood repeatability.

The invention discloses a DNA standard sample for transgenic component detection and application of the DNA standard sample. The DNA standard sample comprises DNA fragments shown as SEQ ID NO.1-7, andthe DNA fragments are specific fragments of components T-E9, T-NOS, P-CaMV35S(P-35S), PAT, T-PinII, P-Rbcs4, T-CaMV35S(T-35S). By detection of the seven targets, most of current commercial transgenicvarieties (at least 63) can be covered, and only a transgenic maize DAS40278 transformant and transgenic soybean DP305423 and CV127 transformants without exogenous components fail to be detected. According to detection, uniformity, stability, definite value and the like of the standard sample all meet requirements in detection, and the standard sample can be applied to quality control in daily detection of corresponding exogenous components, verification and evaluation of detection agents, laboratory proficiency testing activity and the like and is available for commercial popularization andapplication.

The invention discloses a method for detecting paddy rice transgenic ingredients, and belongs to the field of biotechnology. The method comprises: determining a transgenic ingredient, an endogenous standard gene and an exogenous standard gene; preparing a multiple-amplification primer; carrying out sampling and mixing to obtain a mixed sample; extracting a genome of the mixed sample, and adding the exogenous standard gene to obtain mixed nucleic acid; constructing a high-flux sequencing library; carrying out high-flux sequencing on the high-flux sequencing library to obtain a sequencing fragment group; obtaining the number of sequencing fragments of the transgenic ingredient, the number of sequencing fragments of the endogenous standard gene and the number of sequencing fragments of the exogenous standard gene in a to-be-tested sample; according to the number of the sequencing fragments of the exogenous standard gene, determining whether an experiment is successful or not, and calculating the content of the transgenic ingredient in the to-be-tested sample; and according to the content of the transgenic ingredient, determining whether the to-be-tested sample contains the transgenic ingredient. According to the method, various transgenic ingredients in the to-be-tested sample can be quantitatively detected at the same time.

The invention discloses an LAMP (Loop-mediated isothermal amplification) detection primer group and kit and a detection method for a gene vip3A of transgenic plants. The primer group consists of 4 specific primers and has nucleotide sequences shown in SEQ ID NO: 1-4. The detection kit comprises a detection primer solution, DNA polymerase with strand displacement activity, a 10x reaction buffer, a dNTPs solution and a color developing agent. The detection method comprises the steps of carrying out amplification on a DNA template of a sample at the temperature of 63-65 DEG C by adopting the specific primers and the DNA polymerase with strand displacement activity, and judging whether the sample contains a gene vip3A component or not by adopting a color developing agent added color change observing method. The detection primer group and kit and the detection method do not need special instruments, have the characteristics of rapidness, high efficiency, simplicity and convenience in operation, high specificity and the like and are applicable to field detection.

The invention discloses a detection method for detecting exogenous gene components of transgenic salmon in animal products by real-time fluorescence quantitative PCR technology. The method comprises the following steps: firstly, taking DNA of a sample to be detected as a template, performing fluorescent quantitative PCR amplification to obtain PCR amplification products; and secondly, detecting the exogenous gene components of the transgenic salmon by using a real-time fluorescent quantitative PCR technology. 2, detecting fluorescence signals of that amplification product; 3, judging whether that sample contain the foreign gene component of the transgenic salmon according to the Ct value of the detection result; Among them, the reaction system used for PCR amplification contains specific primer pairs and specific probes for amplifying foreign gene components of transgenic salmon. The specific primer pair and probe of the foreign gene component of the transgenic salmon of the inventionare not only good in specificity but also high in sensitivity, which provides a quantitative detection method for quickly and accurately detecting whether the foreign gene component of the transgenicsalmon is contained in the animal products, and has a good application prospect.

The invention discloses a detection method of a tomato transgenic component, which belongs to the field of biotechnology. The method comprises the following steps: determining a transgenic component, an endogenic standard gene and an exogenous standard gene; preparing multiple amplification primers; sampling, and mixing to obtain a mixed sample; sampling a genome of the mixed sample, and adding the exogenous standard gene to obtain mixed nucleic acid; establishing a high-flux sequencing library; performing the high-flux sequencing for the high-flux sequencing library to obtain a sequenced fragment group; obtaining the quantity of the sequencing fragments of the transgenic component in a sample to be detected, the quantity of the sequencing fragments of the endogenic standard gene and the quantity of the sequencing fragments of the exogenous standard gene; according to the quantity of the sequencing fragments of the exogenous standard gene, judging whether the experiment is successful or not, and calculating the content of the transgenic component in the sample to be detected; and according to the content of the transgenic component, judging whether the sample to be detected contains the transgenic component or not. By adopting the method, various transgenic components in the sample to be detected can be simultaneously detected quantitatively.

The present invention relates to an oligonucleotide primer probe for accurate and quantitative detection of the specific gene component of a transgenic rice M12 line, and a kit containing the primer probe. The invention further relates to a digital PCR detection method for quantitatively detecting the specific gene component of a transgenic rice M12 line, wherein the method comprises using the specific oligonucleotide primer and the fluorescent label probe for the specific gene of the transgenic rice M12 line. The present invention further relates to applications of the specific oligonucleotide primer and the fluorescent probe for the specific gene of the transgenic rice line in quantitative detection of the specific gene component of the transgenic rice M12 line. According to the present invention, by using the digital PCR detection method, the content of the specific gene component of the transgenic rice M12 line in the sample can be accurately and sensitively determined, and the sensitivity can achieve 1 copies/[mu]L.