50 results about "Insulin Gene" patented technology

In accordance with the present invention, there are provided humanized fish insulin genes. Humanized insulin the present invention encode human insulin alpha and/or beta chains while using fish-preferred codons and regulatory sequences. These humanized genes are thus expressible in fish islet cells. Also provided are transgenic fish having islet cells containing and capable of expressing humanized insulin genes. These islet cells (Brockmann Bodies) can be xenotransplanted into subjects having diabetes. In this manner normoglycemia can be achieved in the recipient of the islets.

The present invention relates to an adipocyte Insulin adplnsl with Insulin A and B chains of SEQ ID Nos. 1 and 2, a effective method of treating type 2 diabetes in a subject using adipocyte insulin, said method comprising steps of administering the insulin to a diabetic intraperitoneally, an Insulin gene of SEQ ID No. 3, a process of isolating protein Insulin from the adipocytes of the Carp, said method comprising steps of reverse transcripting RNA of adipocytes to obtain cDNA, using oligonucleotide primers of SEQ ID Nos. 5 and 5 to identify AdpInsl gene from cDNA, and deducing amino acid sequence from cDNA to obtain protein Insulin, and an adipocyte of Catla Catla useful for producing the said Insulin.

This invention relates to a method of treating type II diabetes with a polyacetylenic compound of the following formula:

A method for measuring blood levels of β cell DNA that is released upon β cell death by using a quantitative probe technology to detect amplified methylated and demethylated forms of the insulin gene DNA, representing normal tissue and β cell specific origin, respectively. Using probes permits the sensitive and specific identification of demethylated insulin DNA patterns that are present only in β cells. The method offers a bioassay for detecting β cell loss in diabetes, useful for screening of prediabetes, monitoring of disease progression, and selection and monitoring of therapies. The technique finds potential use in both Type I and Type II diabetes, as well as gestational diabetes.

The present invention utilizes transgenic tomato as bioreactor to produce human insulin. The human insulin is produced through designing plant preference codon, artificially synthesizing several DNAsegment, splicing, and adding endoplasmic reticulum residence sequence of KDEL in the C terminal to avoid the degradation of insulin in plant cell. The gene is transformed into tomato via agrobacterium mediating process under the driving of CaMV35S promoter and fruit specificity promoter 2A12 to express human insulin in tomato fruit. This kind of tomato capable of producing human insulin may become one kind of convenient oral product for preventing and treating some diabetes of depending insulin and autoimmune dysfunction, and may be used to extract human insulin for making injection.

The invention relates to the field of genetic engineering and discloses a pair of polypeptides and encoding genes thereof, with sequences shown as SEQ ID NO. 1 to 4. The pair of polypeptides is used for establishing a pair of a recombinant transcription activator like effector (TALE) and a transcription activator like effector nuclease (TALEN); the TALEN is a fusion protein formed by fusing a pair of DNA recognizing proteins with a DNA severing protein respectively; and the TALEN can perform targeted severing on a target site of a human insulin gene so as to achieve targeted modification on the human insulin gene, with the characteristics of strong specificity, high efficiency and high accuracy.

The present invention provides methods for inducing insulin gene expression in cultured pancreas cells, the method comprising contacting a culture of endocrine pancreas cells expressing a PDX-1 gene with a GLP-1 receptor agonist, wherein the cells have been cultured under conditions such that the cells are in contact with other cells in the culture, thereby inducing insulin gene expression in the cells. The invention also provides high throughput screening methods for modulators of β-cell function, stable cultures of cells made by the methods of the invention, and methods of treating a human subject using the methods of the invention.

The invention discloses a method for the in vitro induction of hAMSCs differentiated into ISCs. According to the method, the extracted hAMSCs are cultured for 7 days by a serum-free high sugar-DMEM induction medium containing niacinamide of 10mmol/L and an N2 supplement. The hAMSCs induced by the method of the invention express insulin gene mRNA and PDX-1 gene mRNA, and the content of insulin in the supernatant of the culture reaches above 330muIU/ml; and the method of the invention allows the in vitro induction of the hAMSCs differentiated into the ISCs to be realized.

The invention belongs to the technical field of genetic engineering, and in particular discloses an epinephelus coioides insulin gene, coded protein and an application thereof. The nucleotide sequenceof the insulin protein is shown as SEQ ID NO:1, and the amino acid sequence of the insulin protein coded by the gene is shown as SEQ ID NO.2. The epinephelus coioides insulin gene provided by the invention is obtained for the first time through cloning, and the gene is applied to construction of an expression vector and a recombinant escherichia coli strain; the recombinant epinephelus coioides insulin protein, which is high in expression, is obtained on the basis of the recombinant escherichia coli strain in vitro; and the protein, which can promote glycolipid utilization and growth of suchmarine fishes as epinephelus coioides and the like, can be used for preparing growth promoters or bait additives applicable to the marine fishes; therefore, the protein has a relatively broad application prospect.

The invention relates to a method for detecting specific recombinant human insulin escherichia coli residual host protein, which comprises the steps of taking total host protein prepared by ultrafiltration of escherichia coli fermentation liquid integrating pZRHi-1 plasmid of a human insulin gene as an antigen, stimulating a rabbit to generate an antibody, removing a human insulin antibody by affinity chromatography fixed with human insulin to prepare a specific antibody, labeling the specific antibody with horse radish peroxidase, and then detecting the residual host protein for a sample to be detected by a double antibody sandwich method. The method is high in sensitivity and specificity, and good in repeatability, and can effectively control RHI (recombinant human insulin) product quality.

The invention provides an expression method, a special expression vector, engineering bacteria and an application for recombinant human insulin. According to the expression method, recombinant human insulin genes can be subjected to induced expression in food-grade lactobacillus, and the recombinant human insulin is presented on the surface of the lactobacillus. The engineering bacteria, the expression vector and an inducer, which are used in the expression method, all meet food-grade requirements, so that the potential hazard caused by existence of resistance genes of the expression vector and non-food-grade inducers is avoided. Lactobacillus engineering bacteria containing cell-wall presentation insulin obtained according to the invention can stimulate an NOD (Non-obese diabetic) rat to produce specific antibodies for the recombinant human insulin, so that the level of a cell factor IL-4 related to immune tolerance is obviously increased, and the generation of immune tolerance is induced; a microbial agent can be taken as an oral vaccine for type I diabetes mellitus as long as the induced lactobacillus engineering bacteria are prepared into the microbial agent, complicated post-processing processes, such as purification, are unnecessary, and wide application prospects are obtained.

The present invention is a method for producing an insulin-secreting pre-adipocyte stem cell and the cells derived therefrom. The preferred method comprises obtaining fat cells from a human; treating the fat cells with low-level laser energy; culturing the treated fat cells and suspending them in media together with an insulin gene plasmid; and subjecting the fat cells to transfection. The transfection is facilitated with the application of low-level laser energy.

The present invention discloses one kind of recombinant human insulin and its coding gene and application. The recombinant human insulin is following protein: 1. the protein comprising the amino acid residue sequence of Sequence 1 in the sequence list; and 2. the derivative protein with the sequence of Sequence 1 through the substitution, deletion and/or addition of amino acid residue(s) and possessing the function of human insulin. The recombinant human insulin has a connecting peptide introduced into the chain A and B sequences of natural insulin to favor to forming beta angle structure, a spatial structure similar to that of natural insulin and insulin activity. The present invention also provides one kind of engineering lactic acid bacteria with the recombinant human insulin gene and capable of surviving in intestinal tract for over 48 hr to stimulate the immune system of NOD mouse to generate specific recombinant human insulin antibody, raise the IL-4 level and induce immunologic tolerance generation.

Insulin resistance is a central feature of type II diabetes and other diseases, and may affect every tissue of the body, including the pancreatic beta cell. Insulin signaling is mediated by a complex network of diverging and converging pathways, with alternative proteins and isoforms at almost every step in the process. We have previously shown that insulin activates the transcription of its own gene by signaling through Insulin Receptor A type (Ex11−), PI3 kinase and p70 s6 kinase. When studying the mechanisms underlying the glucose-stimulated activation of the glucokinase gene in pancreatic beta cells, we now demonstrate that also here secreted insulin is a key-factor. In contrast to the insulin gene, transcription of the glucokinase gene is promoted by signaling via Insulin Receptor B type (Ex11+) and protein kinase B (c-Akt). These data provide the first evidence for selectivity in insulin action via the two isoforms of the Insulin Receptor, A type (Ex11-) and B type (Ex11+), and reinforce the concept of the beta cell being an important target of insulin action.

The invention discloses a human insulin gene-containing expression vector and a construction method and application thereof. The plant expression vector pBINOI disclosed by the invention is obtained by inserting a fusion protein expression box which is driven by a rape oil body protein gene promoter and consists of peanut oil body protein genes and human insulin genes between restriction enzyme cutting sites Hind III and Sac I of a plant double expression vector pBI 121. In addition, the invention provides a method for preparing human insulin by converting oil sunflower with the expression vector to construct a plant bioreactor. By adopting the method, the yield of the human insulin can be improved, and the production cost can be greatly reduced; and the method is suitable for industrial production.

β-cell loss in In Type 1 diabetes is typically undetected until the development of hyperglycemia, at which point β-cell mass is significantly reduced. Methylation sensitive quantitative real time PCR (qRTPCR) of demethylated circulating free β-cell specific DNA can be used as a biomarker of β-cell death. Such DNA includes insulin gene and amylin gene DNA. Detection may be by determination of a gene demethylation index. Methylated and demethylated DNA may be distinguished by bisulfite treatment and use of specific PCR primers or probes to detect the different bisulfite treatment products. Detection of demethylated circulating free amylin DNA is useful in identifying β-cell death. The amylin DNA may be used in conjunction with other β-cell specific genes, such as insulin, to provide a multi-gene approach towards the detection of β-cell loss.

Insulin producing β cells are found in three dimensional (3D) structures, the Islet of Langerhans. The 3D structure is required for normal β cell function and survival. β cell pseudoislets (PIs) are useful for study of β cell physiology. Co-culturing of primary human islets and β cell lines together with islet-derived epithelial cells can improve β cell function and survival and maintain the cells' 3D structure, resulting a rapid and spontaneous formation of free-floating PIs. β cells in PIs were similar in size to native islets and showed increased percentage of pro-insulin-positive cells, increased insulin gene expression in response to glucose stimulation, improved glucose-stimulated insulin secretion, and reduced β cell death. Key ECM proteins, absent in monolayer β cells, are deposited by iECs in and round the PIs. iEC induced PIs are a useful tool for examining β-cell/iEC interactions and studying β-cell function in a native 3D configuration.

The invention provides a method for preparing hepatic stem cell decorated by an insulin gene, application of the method to preparation of a diabetes preparation and a preparation comprising the hapatic stem cell decorated by the insulin gene. The invention also discloses the method for preparing the hepatic stem cell decorated by the insulin gene and the application of the method. By the preparation prepared from the hapatic stem cell decorated by the insulin gene, which is provided by the invention, pancreatic island beta cell damaged in the diabetes can be repaired in vivo to secrete insulin so as to provide the possibility of overcoming the defect that the diabetes depends on insulin therapy and radically treating the diabetes.

The invention relates to the field of plant gene engineering and provides a sequence of a human insulin gene designed and synthesized according to plant preferred codons. A plant expression vector isbuilt for the synthetic human insulin gene, so that the gene can be transferred into tomatoes by an agrobacterium rhizogenes-mediated method under the drive of a CaMV 35S promoter and a fruit-specificpromoter 2A12 to express human insulin in tomato fruit. The tomatoes capable of making human insulin can be used as a convenient oral product for preventing and treating insulin-dependent diabetes mellitus and autoimmune disorder diabetes mellitus, and human insulin can be extracted from the tomatoes to be made into injection preparations.

The invention features methods for determining the risk of development of diabetes in a subject by examining the paternal insulin VNTR class. The invention further provides methods to facilitate rational therapy and maintenance of obese patients.

A method for measuring blood levels of DNA that is released upon death from specialized cells in the body, by using PCR or a quantitative probe technology to detect amplified methylated and demethylated forms of cell-specific gene DNA, representing normal tissue and cell specific origin, respectively. Using probes permits the sensitive and specific identification of demethylated cell-specific DNA patterns that are present only in the dying cells. The method offers a bioassay for detecting β cell loss in diabetes based on circulating demethylated insulin gene DNA, and circulating demethylated myelin oligodendrocyte protein (MOG) genes of oligodendrocytes in multiple sclerosis, for example, and may be useful for screening, monitoring of disease progression, and selection and monitoring of therapies.

The invention relates to a microorganism selection culture medium. The culture medium is a liquid culture medium. The culture medium comprises the components: peptone, yeast powder, monopotassium phosphate, disodium hydrogen phosphate, sodium chloride, ammonium chloride, antibiotics and the balance of water. The culture medium is comprehensive in nutrition, simple in preparation and low in cost and has the advantage that the engineered bacterium strain with stability and high expression quantity can be selected. By adopting the culture medium, the selection flow of the recombinant human insulin genetically engineered bacterium high-expression strain is optimized, and an inductive agent does not need to be added in the middle of the strain culture period, so that the selection flow is simplified, and the pollution risk is reduced. Therefore, the culture medium and the preparation method are suitable for the industrialized production of the recombinant human insulin.