99 results about "Transfer gene" patented technology

The invention belongs to the technical field of biology and particularly relates to preparation and cell transfection application of a CS-based (chitosan-based) and CRISPR / Cas9 plasmid-constructed nano gene drug. The preparation comprises dissolving chitosan into acetic acid solution, dropwise adding the chitosan acetic acid solution into water, adding in CRISPR in mass proportion to the chitosan,encapsulating and carrying CRISPR / Cas9 with the chitosan to prepare a novel gene drug nano particle for good gene transfection effects or targeted exogenous gene introduction and further to achieve the effects of transferring gene drugs into cells. The novel nano compound of CRISPR@CS structured on the CS and the CRIPPR through an optimal feeding ratio can significantly improve the transfection efficiency of genes in cells to convey more genes into the cells to take effects, thereby being expected to being widely applied in the field of gene treatment through gene drug delivery.

This invention discloses one EPSPS gene enzyme immune agent case and its use method in the anti-herbicide bean, wherein, the case comprises anti-EPSPS multi-clone antigen, enzyme board with the antigen, enzyme antigens for two, gene switch bean standard product, non-transfer gene bean standard, intense wash liquid, develop liquid and terminal liquid. This invention method comprises the following steps: cloning CP4-EPSPS gene from the gene switch bean expressed in the bacillus coli; then through protein purification complex property to regroup EPSPS protein antibody immune animal to get special single clone antigen and multi-clone antigen; establishing double antigen clamp enzyme immune system test sample to determine its EPSPS protein content.

The invention provides a universal yeast donor strain that contains a conditional centromere and a URA3 allele on every chromosome. This strain was constructed in four rounds of crosses of individual conditional chromosome strains using a novel tetrad-based screen to identify segregants in which all marked chromosomes were contained in the same spore. The invention also provides an improved high efficiency method to transfer extrachromosomal genetic material such as plasmid DNA into any Saccharomyces strain for use with the current gene disruption libraries. The method of transfer is mating-based method which uses a kar1 plasmid donor strain that can initiate mating but cannot form a diploid and allows plasmid transfer (plasmoduction) between nuclei in the heterokaryon. kar1 matings have been used to transfer YACs between yeast strains, but previous methods required specialized genetic backgrounds in the recipient strains and suffered from high rates of spurious chromosome transfer (Hugerat, Y., et al. 1994. Genomics 22:108). Plasmoduction with the universal donor strain only requires that the recipient strain be ura3, GAL+ and have another marker available for selection of the transferred plasmid. Counterselection against every donor chromosome also limits the amount of spurious allele transfer. The universal donor strain and the method of the invention are used to screen the yeast gene disruption library with plasmid-based dominant negative alleles of various genes.

This invention discloses a method for improving the salt resistance and drought resistance of the corn and wheat by aggregating the beta, NHX1, PPase genes with the transfer genes, the steps as follows: regrouping the solution film natrium hydrogen counter-operating protein gene NHX1, the solution film tar phosphatase gene PPase and the Cholinesterase dehydrogenase gene from the escherichia colim to the plant expression carrier, then adding the corn or wheet cells and make them express efficiently, then producing the transfer gene plant; screening out the transfer gene pure cell that has apparently higher salt resistance and drought resistance from the transfer gene plant and its offspring; getting the transfer gene plant with the three object genes through twice tranfering the transfer gene pure cell or making the plant with the different transfer genes mate mutully, then sceening out the ones that have better salt resistance and drought resistance in their offsprings, and making them mute and purify by themselves to creat the new corn and wheat seeds with the salt resistance and drought resistance; or making use of the transfer polymeric material to produce the mixing seeds directly.

This invention relates to a transfer gene soybean checking method and its eduction, which includes:1 distilling soybean DNA;2 PCR expanding reaction, the total volume of the system is 25ul, each component and its final concentration is as follows: cushion solution 10mM, dNTP0.2mMú¼Mg2+1.5mMú¼the up eduction and the down eduction are 0.2mMú¼Taq 1Uú¼DNA50ng respectively, the special eduction react under the condition of the special PCR reaction by filling ddH20 up to the 25ul; after the reaction ending, distills the systerm solution 10ul and makes electrophoresis detection with 2.0úÑ agarose gelatin, and makes record; 3judging whether the soybean is the transfer gene soybean according to the record that whether it can display a 337bp unusual zoster. The related matter includes the up eduction and the down matter, whichare:Fú‘5'-CACTGACGTAAGGGATGA-3 18nt Tmú‘55.0ú¼Rú‘5'-TGTGCTGTAGCCACTGAT-3'18nt Tmú‘55.0.

The invention discloses a method of cultivated silkworm transfer-gene, which comprises the following steps: formulating paste with forced fecula to fix silk seed; utilizing 70% alcohol to sterilize silk seed; arranging the silk seed one by one with fine writing brush; colonizing extra-source gene on the rotor seat carrier; constituting recombinant rotor seat; setting the mass ratio with Helper plasmid as 1 : 1; adjusting the density at 0.4 mu g / mu l with injection buffer liquid; carrying out microscopic injection with stereoscopic microscope; setting the injecting quantity at 15-20ng DNA; sealing with leather and gel; transferring the silk seed and slide into incubation vessel; incubating at 25 deg.c; hatching and seeding; choosing successive; checking.

The invention discloses a cotton salt-tolerant gene, i.e. Gossypium wild dry land cotton CIPK-like protein phosphokinase GarCIPK. The gene has the sequence of SEQ ID No.1 in a sequence list. A cotton stress tolerance related gene is separated by an electronic cloning and reverse transcription-polymerase chain reaction (RT-PCR) technology, and function verification is performed by transferring gene into tobacco and shows that the salt tolerance of a transgenic plant is obviously improved.

The present invention provides a retroviral vector containing a membrane protein having a hemagglutinin activity. The present inventors constructed a retroviral vector pseudotyped by the membrane protein having a hemagglutinin activity. This viral vector showed gene transfer at a high efficiency into host cells. In particular, it was established that genes can be transferred thereby at a high efficiency into cells into which genes can hardly be transferred by the conventional techniques, for example, blood cells and hematopoietic cells including hematopoietic stem cells, and mucous cells including mucosa epithelial cells. The viral vector of the present invention is highly useful as a vector for gene therapy.

This invention discloses a gamma- tocopherol methyl transfer enzyme that from corn and soybean, their code gene and their use in the plant gene project field. This invention uses RACE and RT-PCR technique, it obtains the span cDNA code sequence of gamma- tocopherol methyl transfer enzyme showed by the SEQ ID NO: 1 and SEQ ID NO: 3 from the corn of high oil 115 and soybean (9525) which all belongs to the oil plants, and it obtains the gamma- tocopherol methyl transfer enzyme with biological activity through prokaryotes expression, and it also compares with the difference of gamma-TMT enzyme activity form different sources. This invention transforms the pattern plant from the related code sequence, through HPLC analysis, the content of the alpha- tocopherol of the transfer gene plant leafage is improved by 2-5 times comparing to that of the wild leafage. This invention is to cut a new way for culturing the new plant of oil plants with high vitamin E content.

The invention discloses an alfalfa genetic inverting method in plant transferring gene technical domain, which comprises the following steps: 1, comprising two methods; evoking callus with alfalfa hypocotyls through floating culture mode; assisting by super sound; sieving body embryo in medium; germinating the body embryo; generating root; getting the transforming plant; 2, using tender true leaf of alfalfa as transferring acceptor material; transforming with agricillin medium guild; sieving in traumatic evoked medium; seeding resistance callus in differential medium; evoking out of body embryo; generating root; getting the transforming plant. This invention increases genetic transferring efficiency of alfalfa and shorten genetic transferring period.

The present invention provides for liposomal vectors which inhibit inflammation and / or enhance expression of transferred genes. It is based, at least in part, of the discovery that vectors comprising a nucleic acid carrying a gene of interest and a cationic liposome containing an immunosuppressive agent induce lower levels of inflammatory cytokine relative to conventional lipolex vectors, as well as on the discovery that a component may be incorporated into the liposome which enhances expression of the gene of interest.

The invention relates to a targeting gene transferring method of folic acid-functionalized PAMAM (polyamidoamine dendrimers) wrapped by gold nanoparticles. The targeting gene transferring method comprises the steps of preparing the folic acid-functionalized PAMAM wrapped by gold nanoparticles; preparing a carrier of the folic acid-functionalized PAMAM wrapped by gold nanoparticles and a plasmid compound; and transferring and expressing targeting cervical cancer cells of the compound. The carrier prepared by the invention is used for transferring genes, has the advantages of moderate transferring condition, easiness in operation, high transferring efficiency, good specificity and the like, and has good application prospects in the aspect of cancer treatment and the like.

The invention provides a fluorescent strain E. coli C600 and a construction method and application thereof. By constructing a recombinant plasmid containing an R6k replicon, a transposition unit and an RP4 conjugative transfer gene, the recombinant plasmid is transferred to a host bacterium in which the recombinant plasmid can replicate to obtain a recombinant strain, the recombinant strain and E.coli C600 are subjected to mixed culture, and then the fluorescent strain E. coli C600 can be obtained through screening, wherein the transposition unit is formed by using two ISApl1 for sandwichinga Lux gene cluster and a tellurite drug-resistance gene in the middle. The exogenous gene acquiring capability of the fluorescent strain E. coli C600 is not influenced, the stability is high, stable inheritance can be ensured, the fluorescent property cannot lose along with bacterium passage, and thus the fluorescent strain E. coli C600 can serve as a recipient bacterium for conjugation experiments. By adopting the fluorescent strain E. coli C600, recipient bacteria and zygotes can be identified more efficiently and more intuitively in the fields related to plasmid conjugational transfer, andtherefore the fluorescent strain E. coli C600 has good application prospects.

A bivalent plant expression carrier for culturing high anti-roundup transfer gene plant contains G2-aroA and gat. It has better anti-roundup performance and can be used to culture various high anti-roundup transfer gene plants.

The invention deals with the method of cloning the big critter using the transferring human a-whey albumen gene, the handling process is showed as follows: (1) using whole Human a-whey albumen gene frame to construct the galactophore unusual expressing carrier, (2) constructing the in series fabric connecting the human a-whey albumen unusual expressing carrier and the double single choosing carrier and transmitting the in series fabric DNA to the body nucleolus to gain the transferring gene cell, (3) cloning the body cell using the transmitting gene cell for the nucleus feeding body to gain the transferring clone big critter containing the transferring human a-whey albumen. The transferring gene milch cow galactophore unusually expressing recombining human a-whey albumen can be tapped the health caring drug and the leechdom, the milk containing the recombining human a-whey albumen can be right tapped to the health care milk and the milk produce with the high appending value.

Plumbaginaceae is a highly stress-tolerant angiosperm family consisting of approximately 15 genera of which the genus Limonium is the largest. Many species of Limonium (also called sea lavender and statice) are of commercial importance in ornamental horticulture, especially for cut flowers. It is taught herein that it is possible to generate Limonium plants via somatic embryogenesis and in turn provide a means for repetitive embryogenesis. This repetitive property allows perpetuation of the embryogenic state indefinitely and thereby produce large numbers of Limonium embryos for diverse goals such as mass propagation, production of valuable compounds and for the production of transgenic plants.

A method for converting Chinese ephedra total DNA into yeasts, the transfer gene yeasts thereof, a culture medium for separating the transfer gene yeasts and liquid culture medium for transfer gene yeasts are disclosed.

The invention discloses a method for forecasting genes transferring horizontally in a genome by utilizing a bionic pattern identification principle, which comprises the steps of: utilizing the homologous continuity-based bionic pattern identification principle to forecast the gene transferring horizontally in genome, extracting features of gene sequences, transforming the gene into points in a higher dimensional space, analyzing the point distribution of the same sample in the higher dimensional space, determining a constructing network of a geometric solid covering the sample subspace and forecasting the gene transferring horizontally.

The detecting method of transferring gene product's surface plasma resonate biological sensor, which relates to a detecting method adopts to transferring gene product with surface plasma resonate biological sensor. Its procedure is designing and preparing ssDNA probe modified by tip biotin;using absorption to fix to probe on the surface of chip and forming monomolecular layer to execute detecting chip; adding sampling DNA PCR product to monitor cross reaction and observe analytic result after elution. Leading to the detecting of transferring product,the process is shortcut, accurate and real time response. Not only the psitive result of the determined PCR detected can be verificated, but also the selecting experiment can use the transferring geng product's specificity detecting probe. As compared to general gene detection, the probe is immobilizing, uses repeatedly and is low cost ;the sample needn't deal with in advance and extra reagent, can analyse directly; answering quickly, cutting detecting time and analyzing continuously to make true on line and real time monitoring of DNA; the analysis work is simple and is easy to automatic measurement.

The invention discloses a micro organ culture for separated cell population with special characteristics. The topic micro organ culture has the prominent characteristics of capacity of maintaining a relatively long period in the culture and an organ micro-architecture for facilitating cell-cell and cell-matrix interaction present in a source organ. The micro organ culture can be used in a method for transferring gene products to a receptor subject to identify hyperplasia and cell differentiation reagents and identify and separate progenitor cells from stem cells. In addition, the micro organ culture can be used in a method for identifying hyperplasia, cell differentiation and virus infectious inhibitors. In other schemes, the micro organ culture can be used for transplant.

A long-grained rice classified controlling gene PHR1 and its use are disclosed. It is cloned from long-grained rice 93-11 and approved gene for controlling long-grained classification and named PHR1. Transfer-gene complementary examination proofs that PHR1 is gene for differentiating long-grained rice classification. It can obtain transfer-gene rice with various anti-insect abilities by inhibiting or excess expressing PHR1 to regulate plant resistance by gene engineering technology.

The invention discloses a method for using antibiotic-free selection marker in barbadosnut gene transformation. The method constructs a mannose antibiotic-free selection marker carrier, transfers a phosphomannose isomerase (PMI gene) and a reporter gene (GUS gene) into a barbadosnut cell by mediation by agrobacterium, uses mannose as a selective agent in place of an antibiotic, screens the transformed receptor cell, and detects the transgentic plants by polymerase chain reaction (PCR) to see if the transferred gene is expressed in the cell. Based on the technical scheme provided by the invention, a bud with uncertain resistance is successfully obtained by transforming a barbadosnut leave tray, the genetic transformation rate of cells screened by mannose antibiotic-free selection marker is over 10% which is two times that of cells screened by antibiotics. Besides the high transformation rate, the method also has the characteristics of convenience in detection, high biosafety of transgenic plants, and no potential adverse effect of antibiotics on the environment.

The invention discloses a method for integrating a yarrowia lipolytica genome based on a non-homologous end-linking mechanism. The method comprises the following steps of: integrating yarrowia lipolytica endogenous genes DL4 and XRCC4 and an exogenous wisdom gene PAXX into a genome rDNA site of a yarrowia lipolytica ATCC201249 strain in an rDNA site integration manner, so as to obtain a recombinant strain YNH01; linking a to-be-transferred gene expression cassette with a URA3 nutrition tag U11 truncated to 11bp by a promoter, and transforming an obtained linking fragment into the strain YNH01,so as to obtain a high-pressure screening tag strain; transforming the linking fragment into the high-pressure screening tag strain, so as to obtain a recombinant strain subjected to two-round transformation; and transforming the linking fragment into the recombinant strain subjected to two-round transformation again, so as to obtain a transferred gene high-expression strain. According to the method, a large number of diversified strains are obtained within a short time, and the efficient application of an NHEJ integration technology in microbial synthesis of natural products is realized.

The present invention is aimed to provide oligonucleotide sequence of target activated chronic myeloid leukemia protein kinase PKR and retrovirus dual expression vector possessing the oligonucleotide sequence. The complementary sequence of 20bp SEQ1 and SEQ2 are designed according to BCR-ABL b3a2 type mRNA fusion point upper and down stream, enzyme-cutting point is also designed, a pair of nucleotides T-A is changed to C-G for preventing early stop of transcription.The recombinant retrovirus vector is transformed into K562 cell line of chronic granulocytic leukemia of blast period. The transferred gene is transcribed and hybridizes with BCR-ABL mRNA to form double-chain RNA, activates PKR targetedly, results in K562 cell apoptosis and have no effect on normal cell. The effective ingredient nucleotides sequence is prepared for clinical drug and can treat chronic granulocytic leukemia.

The invention discloses a method to screen and culture barley transfer-gene plant with non-antibiotic of agricillin dielectric conductance in plant transfer-gene technical domain, which comprises the following steps: leading reported gene into receptor barley cell through agricillin dielectric conductance; utilizing mannose to replace antibiotic as chosen agent; screening the reverting receptor cell; adjusting the genetic reforming process of barley through phosphomannose isomerase; further-screening transfer-gene barley with chlorophenol red colouration method and PCR method; getting the end product. The barley cultivating strain conversion rate can reach 12% with agricillin dielectric conductance and can reach 100% with chlorophenol red colouration method.

The invention refers to the ration checking method for transferring gene farm produce and food compound. The method includes folloiwng steps: the extraction of gene group DNA from transferring gene farm produce and food compound; checks the design of sign gene, special gene added ingredient and probe; ascertains the standard curve of ration checking sample; real-time fluorescence ratio polymerase chain reaction (PCR) checking, ration analysis.

The present invention provides methods for producing a viral vector comprising a membrane protein that binds to sialic acid as a component of the envelope, using neuraminidase (NA) derived from Gram-positive bacteria. The methods comprise the steps of culturing cells producing a viral vector in the presence of an NA from Gram-positive bacteria, and recovering the produced virus. The methods of this invention enable the production of high titer virus at high cost performance. Such a viral vector is capable of transferring genes at high efficiency into cells such as blood cells and hematopoietic cells, including hematopoietic stem cells, and mucous cells including mucoepithelial cells, those not amenable to gene transfer by conventional methods, and therefore should be useful as a vector for gene therapy.

This invention discloses a Jatropha curcas slat inducing transcription factor and its coding gene and application. The related enzyme is one the following amino acid remnant radical sequences: 1) SEQ ID íÝ: 1 in the sequence list; 2) the protein that can control the plant resistant reverse and replace, minus or add the acid remnant radical sequences of the SEQ I D íÝ: 1 in the sequence list through one to ten amino acid remnant radicals. Using this transferring gene technique and transplanting the gene in the plant host can obtain the transfer gene plant that has the improved aridity resist and high salt resist ability.

The invention relates to a novel plant genetic transformation method. An ovary injection method is utilized to efficiently and directly introduce vector plasmid DNA dissolved in dimethyl sulfoxide solution into plants so as to obtain transgenic plants.