33 results about "Gene dosage" patented technology

Methods and compositions disclosed herein generally relates to methods of determining minimum hematopoietic stem cell (HSC) chimerism and gene dosage for correction of a hematopoietic disease; in particular, in in vivo models. The invention also relates to modified lentiviral expression vectors for increase a viral titer and various methods for increasing such titers as well as expression vectors capable of enhancing such titers. The invention also relates to CHS4 chromatin insulator-derived functional insulator sequences. The invention further relates to methods for genetic correction of diseases or reducing symptoms thereof, such as sickle cell anemia, a lysosomal storage disease. The invention further relates to a method of improving and / or correcting one or more central nervous system (CNS) abnormalities caused by one or more lysosomal storage disease. The invention further relates to methods of improving titer in transfection-based bioreactor culture production or transfection-based production systems using eukaryotic cells.

The present invention relates to compositions, methods, and kits for quantifying variations in gene copy number, e.g., of individual genes or of chromosomes or portions of chromosomes in an homogeneous reaction, without the need for target amplification, fragment size resolution, or microscopy.

The invention relates to the application of a gene engineering technology in increasing an antibiotic component, in particular to a gene series technology for increasing the main component content ofgene engineering isovaleryl selectomycin. The gene series technology comprises the following steps: connecting ist genes which are closely related to selectomycin isovaleryl acylation in Beta selectomycin gene engineering bacteria in series; increasing the bacterial isovaleryl acylation generating capability by increasing gene dosage; using a promoter erythrocin resistance gene erm promoter sequence with strong promotion activity to replace the original ist gene promoter sequence so as to increase the expression of the series ist genes, and improve the proportion of isovaleryl selectomycin main components of the gene engineering bacteria from a headstream.

The invention discloses an apolipoprotein E4 ELISA kit and a preparation method thereof, the invention applies the purified human apolipoprotein E4 to prepare an anti-human ApoE4 antibody, and an ELISA plate is coated after the compatible purification. The kit composition includes the ELISA plate coated by the anti-human ApoE4 monoclonal antibody, an enzyme-labeled antibody, ApoE4 standard frozen powder, sample dilution solution, TMB substrate color developing solution, concentration washing liquid and reaction termination liquid. The ApoE4 in the antibody capture standard solution or the sample solution which is fixed on the microporous surface of the ELISA plate is identified by and combined with the enzyme-labeled antibody, so as to form the antibody-ApoE4-antibody-enzyme compound, the absorbance is measured after the reaction and color development of the enzyme and the substrate and the concentration of the ApoE4 in the sample can be calculated by the standard curve. The kit can detect the consistency of the apolipoprotein E4 in human serum, plasma or cerebrospinal fluid, the invention has the advantages of sensitivity, rapidness, simpleness and accuracy, so the invention provides the effective means for in vitro diagnosis and determination of ApoE4 gene type and the gene dosage.

The invention relates to a kit for detecting ITD (internal tandem duplication) mutation in an FLT3 (Fms-like tyrosine kinase 3) gene juxtamembrane domain and in particular relates to a kit for detecting the FLT3-ITD in a clinical sample by using a fluorescence PCR (Polymerase Chain Reaction) capillary electrophoresis technology. The kit mainly comprises a PCR amplified reaction solution, a negative control product, a positive control product, a plurality of sealed reagent bottles or pipes which cover the kit as well as packaging boxes which are used for separating and concentratively packaging the reagent bottles or pipes, and the size and gene dosage of each amplified fragment are determined by using an electrophoresis method. The kit disclosed by the invention has high sensitivity and specificity in terms of detection and analysis of the FLT3-ITD in a whole blood or marrow sample and can be widely applied to the fields of judgment prognosis and guide treatment of leukemia and resurgence prevention of diseases.

The invention discloses a selection method of reference genes in quantitative real-time PCR analysis of jerusalem artichoke, and relates to the field of quantitative PCR. The method comprises the following steps: taking different tissues (radicles, immature stems, leaves, stem blocks and petals) of the jerusalem artichoke as materials, and carrying out expression analysis of the reference genes of18S ribosomal RNA gene (18S rRNA), transcription elongation factor gene (Ef-1a), actin gene (Actin and beta-actin), 3-glyceraldehyde phosphate dehydrogenase (GAPDH), 25S ribosomal RNA gene (25S rRNA)and poly-ubiquitin enzyme gene (UBQ)7 by using a q PCR technology; carrying out statistical assessment on the obtained data and analyzing expression change of all housekeeping genes by utilizing GeNorm and NormFinder software, so as to screen out relatively-stable genes as the reference genes of the jerusalem artichoke, which are used for studying the gene dosage changes of the jerusalem artichoke.

Methods and compositions disclosed herein generally relates to methods of determining minimum hematopoietic stem cell (HSC) chimerism and gene dosage for correction of a hematopoietic disease; in particular, in in vivo models. The invention also relates to modified lentiviral expression vectors for increasing a viral titer and various methods for increasing such titers as well as expression vectors capable of enhancing such titers. The invention also relates to CHS4 chromatin insulator-derived functional insulator sequences. The invention also relates to methods for genetic correction of diseases or reducing symptoms thereof, such as sickle cell anemia or β-thalassemia.

Methods and compositions disclosed herein generally relates to methods of determining minimum hematopoietic stem cell (HSC) chimerism and gene dosage for correction of a hematopoietic disease; in particular, in in vivo models. The invention also relates to modified lentiviral expression vectors for increase a viral titer and various methods for increasing such titers as well as expression vectors capable of enhancing such titers. The invention also relates to CHS4 chromatin insulator-derived functional insulator sequences. The invention further relates to methods for genetic correction of diseases or reducing symptoms thereof, such as sickle cell anemia, a lysosomal storage disease. The invention further relates to a method of improving and / or correcting one or more central nervous system (CNS) abnormalities caused by one or more lysosomal storage disease. The invention further relates to methods of improving titer in transfection-based bioreactor culture production or transfection-based production systems using eukaryotic cells.

The invention discloses a breeding method of an erythritol producing strain, belonging to the technical filed of microorganisms. The erythritol producing strain is reconstructed by adopting a genetic engineering means to increase gene dosage of 4-erythrose kinase phosphate and erythrose reductase; and then optimizing a component and a proportion of a culture medium, adding copper chloride in a raw material, optimizing a culture condition, and breeding Yarrowialipolytica strain with lower yield of erythritol so as to obtain a Yarrowialipolytica erythritol high-yield strain. The breeding method specifically comprises the following steps: 1, reconstructing the genetic engineering; 2, preparing a culture medium; and 3, optimizing the culture condition to obtain the high-yield erythritol producing strain. According to the invention, through the reconstruction of the genetic engineering, the blindness of the traditional process is avoided, a large quantity of subsequent screening and detecting works are reduced, the synthetic amount of the 4-erythrose kinase phosphate and erythrose reductase in the Yarrowialipolytica erythritol high-yield strain is greatly increased, and the production of erythritol is facilitated. The breeding efficiency, the strain activity and the yield of the erythritol producing strain are greatly increased.

The invention belongs to the technical field of plant biology, and particularly discloses an echinacea adventitious bud regeneration culture method. DA-6 of different concentrations is added in a regeneration culture medium according to explants of different biomasses and sources. The echinacea adventitious bud regeneration efficiency can be remarkably improved, the problem that the regeneration efficiency of some explants with small biomasses is restrained when DA-6 is added only according to the gene dosage and explant sources originally is avoided, and the purpose of further improving the adventitious bud regeneration efficiency is achieved. The method has wide application prospects and good actual application value, and can generate good economic benefits and social benefits.

The invention discloses a strain for optimizing a fermenting composition of erythromycin, a construction method and application thereof. The strain is saccharopolyspora erythraea which is classified and named Saccharopolyspora erythraea; and the preservation number of the strain is CGMCC2286. Through a means of metabolic engineering, erythromycin is strengthened and is subjected to biosynthesis to modify enzyme; in particular, the strength of C12-hydroxylase of a macrocyclic framework and mycarose methylase; an reconstructed erythromycin promoter is introduced; and through different mixture ratio of gene dosage, the strength and proportion of the hydroxylase and the methylase are adjusted, so as to obtain the strain for only producing a main effective composition of the erythromycin, namely erythromycin A. The strain is adopted to ferment, can reduce impurity compositions, even fully remove the impurity compositions of erythromycin A, erythromycin B and the like, simplify a production process and reduce production cost.

The invention discloses a protease K multi-copy strain construction method, and belongs to the technical field of biology. According to the protease K multi-copy strain construction method, protease Kmulti-copy strains is artificially constructed in vitro by utilizing a Biobrick method, and the protease K multi-copy strain construction method mainly comprises the following specific steps: S1, constructing a multi-copy vector; S2, screening multi-copy strains; and S3, carrying out multi-copy strain large-scale fermentation. A protease K multi-copy tandem expression cassette is artificially constructed in vitro by using a Biobrick method; the construction method has the advantages that the copy number of recombinant plasmids is controllable, dependence on screening of high-concentration resistant drugs is not needed, large-scale screening is also not needed, the gene dose effect is considered as the most important factor for expression of foreign protein by pichia pastoris, and the expression quantity of the multi-copy strains is many times higher than that of single-copy strains; therefore, the yield of the protease K is increased, the price is lower, and the protease K can be applied to various fields.

The invention provides a genetically-engineered strain and a preparation method thereof. According to the invention, a double-auxotroph strain is used as an expression host; two vectors are simultaneously transferred into the expression host through one electrotransformation; one vector contains a target exogenous gene and a compensator gene for one auxotroph of the expression host; and the othervector contains a UPR key gene HAC1p and a compensator gene for the other auxotroph of the expression host. The genetically-engineered strain provided by the invention contains a functional gene and the target exogenous gene of different gene dosages, and high-yield genetically-engineered strain capable of secretory expression of the target exogenous gene can be obtained through screening.

The present invention discloses the preparation process of heat labile enterotoxin of E. coli, and the preparation process has intracellular expression or secretory expression of LT or LTB in yeast cell. Exogenously expressing LT and its mutant or subunit in eukaryotic yeast cell has greatly raised expression level. The present invention constitutes multicopy LT or LTB subunit expression kit, has increased gene dosage of LT or LTB in yeast, LT or LTB gene recombination following alcohol dehydrogenase promoter as the powerful promoter of Pichia yeast expression vector, methanol induced high expression of exogenous protein in yeast cell and thus raised expression level of LT and its mutant or LTB in yeast cell, simple post purification and safe clinical application. The present invention has low production cost and high target protein yield, and is significant in the scale production and clinical application of mucous membrane adjuvant.

Methods and compositions disclosed herein generally relates to methods of determining minimum hematopoietic stem cell (HSC) chimerism and gene dosage for correction of a hematopoietic disease; in particular, in in vivo models. The invention also relates to modified lentiviral expression vectors for increasing a viral titer and various methods for increasing such titers as well as expression vectors capable of enhancing such titers. The invention also relates to CHS4 chromatin insulator-derived functional insulator sequences. The invention also relates to methods for genetic correction of diseases or reducing symptoms thereof, such as sickle cell anemia or β-thalassemia.

A high-throughput assay for characterizing a subject's genetic makeup is disclosed. Specifically, a high-throughput assay utilizing PCR is disclosed that permits the rapid and accurate characterization of a subject's inherited alleles of the polymorphic glutathione S-transferase (GST) genes GSTM1, GSTM3, GSTP1, and GSTT1. This method allows detection of the specific alleles inherited, including the gene dosage of GSTM1 and GSTT1 while not requiring restriction endonuclease digestion of the PCR products in order to detect length differences. Further, the method allows all analyses to be performed simultaneously in the same gel lane, thus further adding efficiency and cost-effectiveness.

The invention belongs to the technical field of plant gene engineering and particularly discloses separation and cloning of alleles Sc-j and Sc-i of a pollen fertility gene loca Sc of paddy rice hybrids, and an application in seed breeding. Sc-j encoding of japonica rice and Sc-i encoding of indica rice contain proteins in a DUF1618 structural domain, which are required in normal growth of pollen of paddy rice. The Sc-j only has one gene copy while the Sc-i, in different varieties, has two or three tandem gene copies. In a hybrid between the japonica rice and indica rice, high-level expression products of the Sc-i multi-copy gene can inhibit expression of the Sc-j, which causes hybrid pollen sterility. The invention discloses a hybridization seed breeding method, in which a part of copies of th Sc-i is mutated by utilizing a gene-editing technology, so that the gene dosage effect of the Sc-i is reduced and further fertility of Sc-j pollen in a japonica-indica paddy rice hybrid is recovered, thus overcoming male sterility of a japonica-indica paddy rice hybrid.

Bacterial live vector vaccines represent a vaccine development strategy that offers exceptional flexibility. In the present invention, genes encoding protective antigens of unrelated bacterial, viral, parasitic, or fungal pathogens are expressed in an attenuated bacterial vaccine strain that delivers these foreign antigens to the immune system, thereby eliciting relevant immune responses. Rather than expressing these antigens using only low copy expression plasmids, expression of foreign proteins is accomplished using both low copy expression plasmids in conjunction with chromosomal integrations within the same live vector. This strategy compensates for the inherent disadvantage of loss of gene dosage (versus exclusive plasmid-based expression) by integrating antigen expression cassettes into multiple chromosomal sites already inactivated in an attenuated vector.

Compositions and methods, systems, and kits for detecting and quantifying variations in numbers of molecules, particularly variations in gene dosage, e.g., due to gene duplication, or to variations from the normal euploid complement of chromosomes, e.g., trisomy of one or more chromosomes that are normally found in diploid pairs, without digital sequencing.

The present invention discloses the preparation process of heat labile enterotoxin of E. coli, and the preparation process has intracellular expression or secretory expression of LT or LTB in yeast cell. Exogenously expressing LT and its mutant or subunit in eukaryotic yeast cell has greatly raised expression level. The present invention constitutes multicopy LT or LTB subunit expression kit, has increased gene dosage of LT or LTB in yeast, LT or LTB gene recombination following alcohol dehydrogenase promoter as the powerful promoter of Pichia yeast expression vector, methanol induced high expression of exogenous protein in yeast cell and thus raised expression level of LT and its mutant or LTB in yeast cell, simple post purification and safe clinical application. The present invention has low production cost and high target protein yield, and is significant in the scale production and clinical application of mucous membrane adjuvant.

The invention relates to the application of a gene engineering technology in increasing an antibiotic component, in particular to a gene series technology for increasing the main component content of gene engineering isovaleryl selectomycin. The gene series technology comprises the following steps: connecting ist genes which are closely related to selectomycin isovaleryl acylation in Beta selectomycin gene engineering bacteria in series; increasing the bacterial isovaleryl acylation generating capability by increasing gene dosage; using a promoter erythrocin resistance gene erm promoter sequence with strong promotion activity to replace the original ist gene promoter sequence so as to increase the expression of the series ist genes, and improve the proportion of isovaleryl selectomycin main components of the gene engineering bacteria from a headstream.

The present invention relates to methods of detecting the presence of a genetic polymorphism within two or more closely linked, homologous genes, for example α-thalassemia, in a sample using RT-PCR by subjecting the sample to separate amplification reactions using (a) a pair of forward and reverse primers specific for the head region of each of said two or more closely linked, homologous genes and (b) a pair of forward and reverse primers specific for the tail region of each of said two or more closely linked, homologous genes; and detecting and quantitating the amplification products relative to a control product.

The invention discloses a strain for optimizing a fermenting composition of erythromycin, a construction method and application thereof. The strain is saccharopolyspora erythraea which is classified and named Saccharopolyspora erythraea; and the preservation number of the strain is CGMCC2286. Through a means of metabolic engineering, erythromycin is strengthened and is subjected to biosynthesis to modify enzyme; in particular, the strength of C12-hydroxylase of a macrocyclic framework and mycarose methylase; an reconstructed erythromycin promoter is introduced; and through different mixture ratio of gene dosage, the strength and proportion of the hydroxylase and the methylase are adjusted, so as to obtain the strain for only producing a main effective composition of the erythromycin, namely erythromycin A. The strain is adopted to ferment, can reduce impurity compositions, even fully remove the impurity compositions of erythromycin A, erythromycin B and the like, simplify a production process and reduce production cost.

Bacterial live vector vaccines represent a vaccine development strategy that offers exceptional flexibility. In the present invention, genes encoding protective antigens of unrelated bacterial, viral, parasitic, or fungal pathogens are expressed in an attenuated bacterial vaccine strain that delivers these foreign antigens to the immune system, thereby eliciting relevant immune responses. Rather than expressing these antigens using only low copy expression plasmids, expression of foreign proteins is accomplished using both low copy expression plasmids in conjunction with chromosomal integrations within the same live vector. This strategy compensates for the inherent disadvantage of loss of gene dosage (versus exclusive plasmid-based expression) by integrating antigen expression cassettes into multiple chromosomal sites already inactivated in an attenuated vector.

The invention discloses the key effector sch9 downstream of the Aureobasidium pullulans TOR pathway and its application. The genome sequence of the effector sch9 is shown in SEQ ID NO.3, the encoded nucleotide sequence is shown in SEQ ID NO.4, and the encoded amino acid The sequence is shown in SEQ ID NO.5; the study of gene function found that after knocking out the sch9 gene, the biomass of Aureobasidium pullulans decreased, and the yield of polymalic acid increased significantly, about 10‑30%; after overexpressing the sch9 gene, The yield of the metabolite polymalic acid decreased, but the yield of exopolysaccharide was significantly increased by about 200‑300%. Therefore, the gene dosage level of the effector sch9 would regulate the conversion of the metabolite anabolism flow, which was beneficial to the increase of Aureobasidium pullulans multi-target Metabolite synthesis is instructive.

Compositions and methods, systems, and kits for detecting and quantifying variations in numbers of molecules, particularly variations in gene dosage, e.g., due to gene duplication, or to variations from the normal euploid complement of chromosomes, e.g., trisomy of one or more chromosomes that are normally found in diploid pairs, without digital sequencing.

The invention relates to the technical field of DNA extraction equipment, in particular to an infant oral cell DNA extraction kit. The kit comprises a connecting pipe and a pipe body movably mounted at one side of the connecting pipe, one side center of the connecting pipe is provided with a connecting hole, one end of the pipe body is inserted in the connecting hole, and the other side center ofthe connecting pipe is provided with a through-hole, pressing plates are rotatably mounted on both connecting pieces through rotating shafts, one ends of the two pressing plates far from the connecting pieces are both equipped with strip-type through channels, connecting plates are movably mounted close to two ends of a spring on a threaded rod, one ends of two tweezers rods far from the connecting plates are fixedly equipped with sampling heads, the corresponding sides of the two sampling heads are connected, and one ends of the two sampling heads far from the tweezers rods are inserted intothe through-hole. The infant oral cell DNA extraction kit provided by the invention has the characteristics of simple structure, convenient operation and easy use, effectively improves the sampling effect, guarantees the sampling quality, and at the same time guarantees the collected gene dosage.

Methods and compositions disclosed herein generally relates to methods of determining minimum hematopoietic stem cell (HSC) chimerism and gene dosage for correction of a hematopoietic disease; in particular, in in vivo models. The invention also relates to modified lentiviral expression vectors for increasing a viral titer and various methods for increasing such titers as well as expression vectors capable of enhancing such titers. The invention also relates to CHS4 chromatin insulator-derived functional insulator sequences. The invention also relates to methods for genetic correction of diseases or reducing symptoms thereof, such as sickle cell anemia or β-thalassemia.