158results about How to "High infection efficiency" patented technology

Adenovirus serotypes differ in their natural tropism. The adenovirus serotypes 2, 4, 5 and 7 all have a natural affiliation towards lung epithelia and other respiratory tissues. In contrast, serotypes 40 and 41 have a natural affiliation towards the gastrointestinal tract. The serotypes described differ in at least capsid proteins (penton-base, hexon), proteins responsible for cell binding (fiber protein), and proteins involved in adenovirus replication. This difference in tropism and capsid protein among serotypes has led to the many research efforts aimed at redirecting the adenovirus tropism by modification of the capsid proteins.

The invention provides two specific T cell receptors targeting G12V or G12C mutation epitopes of a KRAS gene and an anti-tumor application. The two T cell receptors respectively consist of an alpha peptide chain and a beta peptide chain. The invention also provides an antigen binding fragment of the T cell receptors, nucleic acid encoding the antigen binding fragment, a vector containing the nucleic acid, and a host cell containing the vector. The invention further provides a method for preparing a G12V mutation-specific T cell receptor of KRAS or an antigen-binding fragment thereof. The specific T cell receptor and the antigen binding fragment thereof can be used as immune effect activators to stimulate the immune response of the body, thereby generating the action effect of resisting tumors and other diseases.

Retroviral gene therapy vectors that are optimized for erythroid specific expression and treatment of hemoglobinopathic conditions are disclosed.

The invention relates to a recombinant adenovirus vector for efficiently inducing a pluripotent stem cell (PS cell) and method for inducing the PS cell by using the recombinant adenovirus vector. The recombinant adenovirus vector is characterized in that the fiber genes therein are B subgroup adenovirus fiber genes, and an Sox2 gene expression cassette and an Oct4 gene expression cassette are connected within the recombinant adenovirus vector in an operating way. The terminally differentiated cell or the adult stem cell of the mammal, particularly the human is infected in vitro, the Oct4 genes and the Sox2 genes are expressed in an ectopic way, and the terminally differentiated cell or the adult stem cell of the mammal, particularly the human can be induced into the PS cell efficiently and rapidly under the synergistic effect of adjusting the epigenetic-inheritance small-molecular medicament. The PS cell of the human can be used for the cell replacement therapy to treat diseases, and the PS cell of the mammal can be used for preparing the transgenic animal model and the animal disease model.

The invention provides a lentivirus recombinant expression vector/a recombinant lentivirus, as well as application, a host cell and a preparing method thereof. The encoding gene of an estrogen related receptor (ERR) protein can be inserted in a host gene group by the lentivirus recombinant expression vector/the recombinant lentivirus which is provided by the invention through a gene recombinant so as to continuously and stably express the ERR. The ERR can be permanently and stably expressed by the host cell which is provided by the invention, and sufficient high-quality virus liquid can be provided for animal in-vivo experiments. Besides, the lentivirus recombinant expression vector/the recombinant lentivirus which is provided by the invention has a wide host range, can infect various cells, such as nerve cells, muscle cells, liver cells, tumor cells and endothelial cells and has a very wide application range.

The invention discloses spore suspension for improving the yield of white muscardine silkworms and application of the spore suspension. The spore suspension for improving the yield of the white muscardine silkworms comprises the following raw materials: muscardine spore powder with 1x106-5x106 muscardine spores per mL, 0.2 part by mass of polysorbate-80, 1-3 parts by mass of cane sugar, 0.4-0.6 part by mass of peptone, 0.05 part of auxiliary materials and the balanced sterile water. The auxiliary materials can be magnesium sulfate, chloramphenicol and streptomycin sulfate. By the spore suspension for improving the yield of the white muscardine silkworms, the concentration of the optimized muscardine suspension is increased, the stickiness and the activity of the muscardine suspension are improved, the germination rate of the muscardine suspension is increased, and the yield and the quality of the white muscardine silkworms are improved obviously.

The invention discloses an efficient, rapid and stable genetic transformation method for strawberries 'beauty'. The method comprises the steps of activation of culture, preparation of plant materials, infection, co-culture, screening culture and rooting culture, solves the problems of the pollution of agrobacterium and infectious microbes in a traditional agrobacterium-mediated method, low conversion efficiency of the strawberries 'beauty' and long period of the strawberries 'beauty' from transgenosis and transgenic detection to field transplanting, shortens the period, further solves the problems of large workload and heavy work of transgenic detection, simplifies a detection method, and increases the detection speed. On the basis of the established 'beauty' strawberry leaf regeneration system, the method researches various factors for influencing agrobacterium tumefaciens transforming the 'beauty' strawberry leaves, optimizes various parameters, simplifies the operation process, and determines the best transformation conditions.

The present invention provides a recombinant glandular related virus capable of expressing human kallikrein, its preparation method and application, and medicine composition containing said recombinant glandular related virus. Said recombinant glandular related virus can drive the human kallikrein gene to make long-period expression in human body, and possesses good physiological and pathologic physiological action for effectively reducing blood pressure, improving renal function and preventing apoplexy, and can be used for clinical curing the above-mentioned diseases.

The invention discloses an efficient genetic transformation method of vacuum permeation assistance soybean immature cotyledon regeneration system, relating to the technical field of plant genetic engineering, comprising the following steps: (a) the induction and proliferation of soybean immature cotyledon somatic embryo; (b) the vacuum permeation assistance introduction of foreign DNA; (c) the regeneration of the somatic embryo with foreign DNA into a plant. Adopting the vacuum permeation assistance agrobacterium mediating method, under a certain vacuum condition, after treat for a certain time, a plurality of micromechanical wounds are made on the soybean somatic embryo, which fully raises the intruding opportunity for agrobacterium to obviously raise the infection efficiency of agrobacterium and the conversion of the soybean compared with the common agrobacterium mediating method. Soybean immature cotyledon is used as an explant to transform genetically, which avoids the problem that chimerism easily, appears with the cotyledonary node as the explant.

The invention discloses an inducing method for danio-rerio inductivity multi-potential stem cells. The induction method includes the following steps that danio-rerio tissues or danio-rerio embryos are gotten, and danio-rerio fibroblasts are obtained through in-vitro isolated culturing; the danio-rerio fibroblasts are infected through viruses, the infected danio-rerio fibroblasts are induced, and the danio-rerio inductivity multi-potential stem cells are obtained through manual selecting cloning and subculturing. Based on the same technical scheme, the invention further discloses an inductive culture medium and an iPS culture medium used for the inducing method. RT-PCR analysis indicates that iPS cells induced through the embryo fibroblasts have the potency of being differentiated into various triploblastic-source cells. By means of the method, the inductive culture medium and the iPS culture medium, the technical difficulty that the infection efficiency of slow viruses in fish cells is low is overcome, and new tool cell sources and new concepts are provided for researching a danio-rerio somatic-cell reprogramming mechanism to select cell materials.

The invention provides a chimeric antigen receptor T cell and application thereof. The chimeric antigen receptor T cell expresses chimeric antigen receptors targeting CD19 and PD-L1 at the same time.The chimeric antigen receptor T cell expresses a CD19 CAR structure and a PD-L1 CAR structure at the same time, and after PD-L1 CAR is combined with PD-L1 expressed by a tumor cell, an activation signal can be transmitted intracellularly. Therefore, the chimeric antigen receptor T cell not only can recognize the tumor cells expressing CD19 in a targeted manner, but also can relieve immunosuppression of PD-1 expressed by the tumor cells on the T cells, avoid an immune escape mechanism of the tumor cells and relieve the problem of drug resistance of CAR-T treatment; and a new idea is provided for CAR-T treatment and adoptive feedback immune cell treatment.

The invention provides an oncolytic adenovirus vector for modifying and expressing two exogenous genes by fibrin. The oncolytic adenovirus vector provided by the invention is characterized in that: a human adenovirus 5 type gene group has a lacking of 24bp basic group in 922bp-947bp; and an expression element for expressing a first exogenous gene is inserted into 28183bp-29906bp of the human adenovirus 5 type gene group, a fibrin chimera is inserted into 31042bp of the human adenovirus 5 type gene group and an expressing element for doubly expressing a second exogenous gene and an eGFP (Green Fluorescent Protein) is inserted into 32021bp and 32022bp of the human adenovirus 5 type gene group. A construction method of the oncolytic adenovirus vector provided by the invention comprises the following steps of: constructing a shuttle vector lacking pAd5 E1A 24bp, a pHBD24 adenovirus vector framework, a shuttle vector of a pHBDE3-first exogenous gene, a pHBDE3-first exogenous gene/SwaI condition copied adenovirus vector framework and a pshuttle Ad5-E4-fibrin chimera shuttle vector; expressing a shuttle vector/ second exogenous gene-E4-fibrin embedding body sequence of the eGFP and the second exogenous gene; and preparing the oncolytic adenovirus vector for modifying and expressing two exogenous genes by fibrin. The vector provided by the invention can be used for inhibiting malignant glioma, liver cancer, stomach cancer, colon cancer, breast cancer and melanoma.

The invention belongs to the biological field and particularly relates to preparation and application of recombinant porcine circovirus PCV2-Cap/Phage display particle. A preparation method of the capsid protein phage display particle of recombinant II porcine circovirus specifically comprises the following four steps: (1), optimizing and coding nucleotide of the capsid protein of the recombinant II porcine circovirus; (2), constructing a recombinant plasmid; (3), constructing a recombinant phage genome; and (4), preparing the capsid protein phage display particle of recombinant II porcine circovirus. An animal experiment proves that the recombinan display plasmid has good antigenicity, can stimulate a pig body to generate good humoral immunity and cellular immunity reaction, and can prevent and control II porcine circovirus infection; time needed for copying the process is short, production cost of the recombinant display particle is low, and product is convenient to use, simple and convenient to store and high in cloning yield.

The invention discloses a dynein mosaic type recombinant human type-B adenovirus and a preparation method thereof. The skeleton of the dynein mosaic type recombinant human type-B adenovirus is a human type-B adenovirus genome, and a base sequence which encodes a receptor binding domain of dynein is a base sequence which encodes a corresponding domain of a human type-C adenovirus. By a molecular cloning method, Ad5-knob gene fragments are cloned and replaced to recombinant shuttle plasmids, in-vitro recombinant on the Ad5-knob gene fragments and a recombinant human type-3 adenovirus genome is realized, obtained knob gene fragments are replaced into type-5 recombinant human type-3 adenovirus genome, and therefore, dynein mosaic type recombinant human type-3 adenovirus rAd3-FK5 is obtained. The dynein mosaic type recombinant human type-3 adenovirus rAd3-FK5 can be infected with mouse primitive epithelial cells and golden hamster lung and kidney primitive cells in vitro, and the infection efficiency of the dynein mosaic type recombinant human type-3 adenovirus rAd3-FK5 is close to that of Ad5, and is much higher than that of a parent strain rAd3E, in golden hamster cells, significant copying exists, and the dynein mosaic type recombinant human type-B adenovirus can be used for small animal model research of human type-3 adenovirus vaccines and antiviral drug evaluation.

The invention discloses isolation and identification of Yunnan tomato leaf curl viral genome and agrobacterium tumefaciens-mediated infective clone construction thereof. Total DNA of a genome of an infected tomato is extracted, virus total genomic DNA is cloned by using PCR (Polymerase Chain Reaction), and a total genomic sequence of a Yunnan tomato leaf curl virus is obtained through sequence measurement. 1.4 direct repeated genomes are obtained through methods of PCR and restriction enzyme digestion and are inserted into a plant expression carrier pBinPLUS with high replication capacity, a recombinant carrier is imported with agrobacterium tumefaciens strains EHA105 with strong infection capacity through an electroporation method, therefore, the agrobacterium tumefaciens-mediated infective clone has high-efficiency infection capacity to a host plant. The invention provides a matured method and system for researching interaction among the host plant, a virus infecting medium and a virus, field detection, genome structure and function research of the Yunnan tomato leaf curl virus.

The invention relates to a genetic engineering technology, in particular to a recombinant virus vector. The vector includes the virus vector and at least two nucleotide sequences, the nucleotide sequences respectively code the antigens of pathogenic virus, the antigen comprises a B cell epitope and a T cell epitope; the recombinant virus vector can lead immune animals to produce humoral immune and cell immune reactions, thus achieving the purpose of the prevention and treatment of diseases. The injection of the recombinant adenovirus vector of the invention has high safety at the same time.

The invention relates to the technical field of animal vaccines, and especially relates to a rescue method for a H9N2 subtype cold-adapted live attenuated vaccine transfected with 293T and DF-1 co-cultured cells. A H9N2 subtype cold-adapted attenuated strain TX-25-CE30 is selected to construct 8 plasmids expressing the gene, the internal gene of TX-25-CE30 is used as a skeleton, the parental strain TX strain HA and NA genes are taken as external genes, the human embryonic kidney cell 293T and chicken fibroblast cell line DF-1 co-cultured cells are transfected, the strain that has weaker ability to infect mammalian cells is rescued and named as rTXca-HA-NA, and the immune protection rate of the recombinant virus after immunization of the chicken to a homologous strain can reach 100%.