36 results about "Beta globin gene" patented technology

The invention relates to a method for detecting human beta-globin gene mutation, in particular to a method for detecting the beta-globin gene mutation by a modified molecular beacon melting curve. The method comprises the following steps of: designing and preparing a corresponding modified molecular beacon in region of a beta-globin gene needing mutation detection; designing an upstream primer and a downstream primer on the periphery of the designed modified molecule beacon, and performing PCR amplification on fragments containing the region to be detected by using the upstream primer and the downstream primer; and after PCR amplification is finished, analyzing the melting curve, and judging whether a nucleotide sequence to be detected has the gene mutation and possible mutation types according to the change of the melting point of the modified molecular beacon.

The invention discloses a recombinant expression vector for stably and efficiently expressing a human beta-NGF (Nerve Growth Factor), a recombinant cell strain containing the recombinant expression vector and a method for producing a recombinant human beta-NGF. The recombinant expression vector comprises a promoter, a beta globin gene intron and a human beta-NGF gene, an internal ribosome entry site sequence and a selective marker gene and has higher integration and expression efficiency in a host cell. A recombinant eukaryotic cell containing the recombinant expression vector expresses the recombinant human beta-NGF equivalent with a natural beta-NGF in activity and still can stably express after multiple times of passage; and the expressed beta-NGF is secreted into a culture medium and is easy to separate and purify, the production downstream difficulty and the production downstream cost are greatly reduced, and the clinical application prospect and the commercial value are good.

The invention discloses a nucleic acid detection kit for 12+2 high-risk human papilloma virus (HPV), comprising: (1) probes as shown in SEQ ID No:1-12 and primers as shown in SEQ ID No: 13-36 of a nucleotide sequence which is only complementary with the DNAs of 12 high-risk HPV; (2) a probe as shown in SEQ ID No:37 and primers as shown in SEQ ID No: 38-39 of a nucleotide sequence which is only complementary with the DNA of high-risk HPV16; (3) a probe as shown in SEQ ID No:40 and primers as shown in SEQ ID No: 41-42 of a nucleotide sequence which is only complementary with the DNA of high-risk HPV 18; and (4) a probe as shown in SEQ ID No: 43 and primers as shown in SEQ ID No: 44-45 of a nucleotide sequence which is only complementary with the DNA of an intracellular beta-globin gene, wherein each group of probes respectively mark different dyes with the maximum emission wavelengths of 518nm, 538-553nm, 574-575nm, 607-615nm, 640nm, 666-667nm and 690nm. The kit disclosed by the invention can furthest lighten huge cost brought by HPV gene detection in China, and the health of people is protected.

The invention relates to a fluorescence quantitative PCR detection kit for beta-thalassemia mutation. The kit comprises a PCR mixing reaction liquid, a positive control and fluorescence probes for detecting beta-thalassemia mutation genotype, wherein the PCR mixing reaction liquid contains PCR primers for amplifying a gene fragment on which a mutation site is positioned, and the mutation site is at least one mutation site selected from deletion mutation of a base corresponding to the site 41 / 42 amino acid of beta-globin gene, C-to-T mutation of a base corresponding to the site 654 amino acid of the second intron of beta-globin gene, A-to-T mutation of a base corresponding to the site 17 amino acid of beta-globin gene, A-to-G mutation of a base corresponding to the site 28 amino acid on the upstream of the promoter of beta-globin gene, A base insertion mutation of a base corresponding to the site 71 / 72 amino acid of beta-globin gene, and G-to-C mutation of a base corresponding to the site 5 amino acid of the first intron of beta-globin gene. With the technical scheme of the present invention, rapid, accurate and high sensitivity detection of mutation conditions of beta-thalassemia gene can be achieved, and especially 6 special site mutations of beta-thalassemia gene can be detected, wherein the 6 special site mutations are common in Chinese.

Methods of using one or more fumaric acid esters or pharmacologically active salts, derivatives, analogues, or prodrugs thereof to increase expression of fetal hemoglobin (HbF) are disclosed. The methods typically include administering to a subject an effective amount of one or more fumaric acid esters optionally in combination or alternation with hydroxyurea to induce HbF expression in the subject in an effective amount to reduce one or more symptoms of a sickle cell disorder, a hemoglobinopathy, or a beta-thalassemia, or to compensate for a genetic mutation is the human beta-globin gene (HBB) or an expression control sequence thereof. Pharmaceutical dosage units and dosage regimes for use in the disclosed methods are also provided.

The invention provides a human papillomavirus gene chip, a preparation method and application thereof, an assay kit and application thereof. The gene chip comprises a solid-phase vector and an oligonucleotide probe fixed on the solid-phase vector, wherein the oligonucleotide probe contains a DNA fragment or a complementary DNA fragment thereof selected from L1 genes of the human papillomavirus or human beta globin gene. The invention also provides a preparation method for the gene chip and an assay kit containing the gene chip. The gene chip and the kit have the characteristics of simple and convenient operation, high flux, high accuracy, strong repeatability and the like, can achieve the aim of detecting 25 different types of human papillomavirus, and can be applied to the clinical test, epidemiology analysis and the like of medical and health departments.

A method is described for regulating gene expression related to iron metabolism to ameliorate diseases that include sickle cell disease, cancers, neurodegenerative diseases, Friedreich's ataxia and other neuromuscular disorders, and atherosclerosis. This approach is illustrated by recent findings that show that ferritin-H, an iron-binding protein that is present in cell nuclei, can repress the human beta-globin gene, the gene that is mutated in sickle cell disease. Increased expression of ferritin-H or a related ferritin-family peptide, given to effected cells either as the peptide itself (or a part thereof), as an expression clone of the ferritin-H-subfamily gene, or via a gene regulator that increases expression of the ferritin-H-subfamily gene itself, prevents or ameliorates expression of the disease state in disorders where increased availability of iron is implicated in the etiology of the disease, including those named above.

The invention relates to a thalassemia-induced multipotent stem cell as well as a preparation method and application thereof. The multipotent stem cell is derived from a body cell of a thalassemia patient; and a genome of the thalassemia-induced multipotent stem cell contains a normal beta globin gene. The thalassemia-induced multipotent stem cell disclosed by the invention can be differentiated into hemopoietic stem cells under proper conditions; and the hemopoietic stem cells can express the normal beta globin gene.

The invention relates to the biomedical field, in particular to a parkinsonism mouse model, a method for establishing the parkinsonism mouse model and application of the parkinsonism mouse model. The invention transfers an extrinsic BAG5-Flag expression cassette to a genome of the animal model, and the expression cassette comprises: (a) a promoter of the rat tyrosine hydroxylase, (b) a part of Beta-globin gene and BAG5-Flag, and (c) a human somatotropin mini gene. The invention also provides the method for establishing the parkinsonism mouse model, and the parkinsonism mouse model obtained by the method of the invention can be used for studying an ubiquitin-protein degradation system and a pathology process mechanism, and screen medicine applied to delay and treatment of parkinsonism.

The invention discloses a target sequence complementary quenching probe and a kit for detecting beta globin gene point mutation. The kit comprises an asymmetric polymerase chain reaction (PCR) single-stranded deoxyribonucleic acid (DNA) template enrichment PCR amplification primer, the target sequence complementary quenching probe, LA DNA polymerase, and the like. A beta-thalassemia point mutation genotype of a tested sample can be detected by using an asymmetrical specific PCR single-stranded DNA template to enrich a human beta-globin gene mutation site target sequence and using a specific fluorescent probe to perform molecular hybridization and melting analysis. The kit has good sensitivity and accuracy for detecting wild type and mutant type of a beta-thalassemia mutation site, is excellent in repeatability and stability, and is completely suitable for the clinical detection of beta-thalassemia point mutation.

The invention discloses a mycoplasma pneumoniae detection method based on a PCR-internal control nucleic acid test strip. The method specifically comprises the following steps that an MP specific primer modified with digoxin and biotin is designed according to the 16S rRNA nucleic acid sequence of mycoplasma pneumoniae, and a beta globin gene primer modified with digoxin and fluorescein is designed according to the nucleic acid sequence of a human beta globin gene; the DNA of a to-be-detected sample is extracted and serves as a template, the MP specific primer and the beta globin gene primer are used for PCR amplification to obtain an amplified product of the DNA of the to-be-detected sample, and the UNG enzyme anti-pollution technology and internal control are added for avoiding false positive and false negative results respectively. The amplified product of the DNA of the to-be-detected sample is detected through the mycoplasma pneumoniae detection internal control nucleic acid teststrip. The mycoplasma pneumoniae detection method based on the PCR-internal control nucleic acid test strip is easy to operate and capable of quickly detecting mycoplasma pneumoniae.

The invention discloses a lentiviral vector suitable for gene therapy of thalassemia and sickle anemia. The system comprises: a, a micro locus control region, which is a micro regulatory element which is screened from a locus control region of beta globin 16kb and does not contain an HS1 region; b, a gene sequence of beta globin, wherein the gene sequence is as shown in SEQ ID NO.3; c, a promoter sequence of an upstream flanking of the gene sequence of beta globin; d, a flanking sequence at the downstream of the gene sequence of beta globin; and e, an insulator sequence from a Foamy virus (Foamy virus). Compared with the prior art, the invention optimizes the structure of the vector, and finds that after the second intron of the Beta globin gene (HBB) is optimized, the expression of the gene can be improved to a certain extent, and the packaging yield of the virus can be greatly improved, so that the clinical use and industrialization of the Beta globin gene become possible.

An objective of the present invention is to provide a microarray for detection of mutations in beta-globin genes with which a large number of mutations (subject for detection) can be simply and quickly detected. The present invention provides a probe group for detection of mutations in beta-globin genes including genes represented by SEQ ID NOs: 3, 4, 7, 8, 11, 12, 17, 18 and 25 to 66, a microarray in which the probe group is immobilized, a method for detection of mutations in beta-globin genes using the microarray, and a kit for detection of mutations in beta-globin genes using the microarray and primers.

Provided herein are adeno-associated virus (AAV) compositions for correcting a mutation in a beta globin gene (HBB) gene and methods of using the same to correct an HBB gene mutation in a cell. Also provided are packaging systems for making the adeno-associated virus compositions.

The invention discloses a nucleic acid composition for detecting HPVs (Human Papilloma Viruses) as well as application and a kit of the nucleic acid composition, and relates to the technical field ofin-vitro diagnosis of virus nucleic acid detection. The nucleic acid composition for detecting the HPV comprises primers which are shown in SEQ ID NO.1 to 6 and primers which are shown in SEQ ID NO.7to 9. The nucleic acid composition disclosed by the invention is capable of simultaneously detecting HPV 16 and HPV 18 and parting the specificity of the HPV 16 and the HPV 18, meanwhile, quality control can also be carried out on a detecting process through an internal control beta-globin gene detection result, false negatives can be reduced, and the nucleic acid composition has the characteristics of high sensitivity and strong specificity.

The invention discloses a nucleic acid detection kit for 12+2 high-risk human papilloma virus (HPV), comprising: (1) probes as shown in SEQ ID No:1-12 and primers as shown in SEQ ID No: 13-36 of a nucleotide sequence which is only complementary with the DNAs of 12 high-risk HPV; (2) a probe as shown in SEQ ID No:37 and primers as shown in SEQ ID No: 38-39 of a nucleotide sequence which is only complementary with the DNA of high-risk HPV16; (3) a probe as shown in SEQ ID No:40 and primers as shown in SEQ ID No: 41-42 of a nucleotide sequence which is only complementary with the DNA of high-risk HPV 18; and (4) a probe as shown in SEQ ID No: 43 and primers as shown in SEQ ID No: 44-45 of a nucleotide sequence which is only complementary with the DNA of an intracellular beta-globin gene, wherein each group of probes respectively mark different dyes with the maximum emission wavelengths of 518nm, 538-553nm, 574-575nm, 607-615nm, 640nm, 666-667nm and 690nm. The kit disclosed by the invention can furthest lighten huge cost brought by HPV gene detection in China, and the health of people is protected.

The invention relates to the field of biomedical treatment, in particular to a gene expression cassette, a lentiviral vector and application of the lentiviral vector in treatment of beta thalassemia. The gene expression cassette comprises a promoter, a beta globin gene, an intron-BCL11A-shRNAmir and a polyadenylation signal which are sequentially connected; and the promoter is a type II promoter with erythroid cell specificity. The gene expression cassette can specifically up-regulate expression of beta globin and gamma globin in erythroid cells, and the two mechanisms have a synergistic effect, so that the treatment effect is remarkably enhanced.

A nucleic acid hybridization membrane strip used for diagnosing the beta-Mediterranean anema is composed of a substrate and the specific probes fixed to the substrate. Said each specific probe (mutant probe) is relative to one of the mutant beta-globin gene sites and has the sequence of 14-20 bases. Its reagent kit is also disclosed.

Methods of using one or more fumaric acid esters or pharmacologically active salts, derivatives, analogues, or prodrugs thereof to increase expression of fetal hemoglobin (HbF) are disclosed. The methods typically include administering to a subject an effective amount of one or more fumaric acid esters optionally in combination or alternation with hydroxyurea to induce HbF expression in the subject in an effective amount to reduce one or more symptoms of a sickle cell disorder, a hemoglobinopathy, or a beta-thalassemia, or to compensate for a genetic mutation is the human beta-globin gene (HBB) or an expression control sequence thereof. Pharmaceutical dosage units and dosage regimes for use in the disclosed methods are also provided.

The invention relates to a thalassemia-induced multipotent stem cell as well as a preparation method and application thereof. The multipotent stem cell is derived from a body cell of a thalassemia patient; and a genome of the thalassemia-induced multipotent stem cell contains a normal beta globin gene. The thalassemia-induced multipotent stem cell disclosed by the invention can be differentiated into hemopoietic stem cells under proper conditions; and the hemopoietic stem cells can express the normal beta globin gene.

The invention discloses a human-source chromosome targeting carrier and preparing method and appliance of beta-globin gene cluster, which is characterized by the following: the carrier contains recombinant human-source chromosome targeting BAC carrier of Beta-globin gene cluster,BGGC, whose GenBank number is NG_000007 to construct carrier as human-source chromosome targeting BAC carrier BACMS.

The invention discloses a kit for detecting α and β globin gene sequences in one tube, including amplification primers, and used for identifying α globin gene sequences -SEA, -α 3.7 ,-α 4.2 Site deletions, alpha globin gene sequence CD30, CD31, CD59, WM, QS, CS site mutations and beta globin gene sequence ‑82, ‑80, ‑79, ‑78, Cap+1, Cap40‑43, Int , CDs14 / 15, CD17, CD19, CD26, CDs27 / 28, CD30, IVSⅠ‑1, CD31, CD37, CDs41 / 42, CD43, CDs71 / 72, CD95, IVSⅡ‑5, IVSⅡ‑654 site mutation probe .

The invention discloses deletion beta thalassemia detection primers and a beta thalassemia detection kit based on SNP analysis and application of the deletion beta thalassemia detection kit. The sequences of the detection primers are shown as SEQ ID NO.1-14. The detection kit provided by the invention comprises the primers. The detection primers refer to six SNP loci, after a to-be-detected sampleis subjected to PCR amplification, whether beta-globin gene cluster deletion mutation occurs or not is judged according to whether heterozygous types of the sample SNP genotypes exist or not. If theexperiment result shows that heterozygous loci exist in 6 SNPs of the sample, nine kinds of beta-globin gene cluster deletion mutation can be excluded. Otherwise, if the typing detection results of the six SNP loci are all homozygotic types, the sample is quite likely to be subjected to beta-globin gene cluster deletion mutation, resulting in loss of heterozygosity, and it is prompted that furtherclinical detection needs to be conducted. Typing detection to the SNPs by the detection primer and kit is easy and convenient to operate, and the cost is low.

The invention provides an erythroid-specific human beta globin gene promoter, a recombinant sequence for expressing human beta globin and applications thereof, belonging to the technical field of genetic engineering. The present invention provides an erythroid-specific human β-globin gene promoter, the nucleotide sequence of which is shown in SEQ ID NO: 1, which has the characteristics of high-efficiency and erythroid-specific activation of functional gene expression. The present invention also provides a recombinant sequence specifically expressing human β-globin in erythroid cells, which is human β-globin gene locus control region HS3~1-β-globin gene promoter-β-globin gene-β-globin Gene enhancers. By constructing a recombinant vector expressing the recombinant sequence, it was verified in erythroid cells that the recombinant sequence can specifically express the human β-globin gene with high efficiency, which provides a basis and experimental reference for gene therapy of β-thalassemia, and is useful for the treatment of thalassemia. It is of great significance, and at the same time provides method support for the study of the expression regulation of β globin.

The invention discloses a method for preparing a beta<IVS-II-654> thalassemia transgenic mouse model carrying human beta-globin gene. A lentiviral vector mediated human beta-globin gene is stably integrated in the genome of the mouse model, and the genome can be expressed continuously and stably; furthermore, the genome can also be inherited to the offspring and maintain the original expression level, thus phenotypes such as hematology, pathology and the like of the beta<IVS-II-654> thalassemia transgenic mouse model carrying the human beta-globin gene are improved. The mouse model prepared by the method can be used for evaluating the effectiveness, the stability and the security of the lentiviral vector mediated human beta-globin gene in the treatment of beta<IVS-II-654> thalassemia.

The embodiment of the invention relates to the technical field of biological detection, in particular to an anemia screening kit for capturing long-fragment DNA in a targeted mode through CRISPR and CAS9. The anemia screening kit comprises sgRNA used for capturing thalassemia related genes in a targeted mode, a Y-type connector used for cyclization amplification and a primer used for cyclization amplification. The target capture range of the sgRNA is as follows: chr11: 5166361 to 5296578 (hg38), and chr16: 48994 to 210817 (hg38), and the target capture range of the sgRNA is as follows: chr11: 5166361 to 5296578 (hg38). According to the anemia screening kit for target capture of long fragment DNA by CRISPR and CAS9 and the method thereof, a large fragment region of thalassemia variation is captured in a targeted manner through CRISPR and CAS9 systems, five types of thalassemia mutation regions including alpha, beta, delta, delta beta and epsilon gamma delta beta are covered, and a three-generation or four-generation long-reading sequencing technology is utilized to identify the thalassemia mutation region of the thalassemia mutation region of the thalassemia mutation region of the thalassemia mutation region of the thalassemia mutation region of the thalassemia mutation region. The kit can effectively detect pathogenic mutation, complex structure variation and large fragment deletion which may cause thalassemia in people, and non-deletion and deletion mutation covering beta globin genes and related regulatory genes including HS-40 and beta LCR genes.