Transformant for reducing weight and fat and preparation method thereof
A transformant, oxyntomodulin technology, applied in the field of biomedicine, can solve the problems of poor acceptance by obese patients, short half-life in the body, and high production costs, and achieve the effects of overcoming high production costs, improving curative effect, and low cost
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
example 1
[0032] The preparation of example 1OXM gene transformation bifidobacterium
[0033] 1. Construction method of OXM Bifidobacterium-Escherichia coli recombinant expression plasmid:
[0034] 1.1 Chemical synthesis of extracellular exo-xylanase (exo-xylanase) secretory signal peptide nucleic acid sequence Xs (SEQ NO.1); using pBAD / gIII plasmid DNA as a template with primers 1 (SEQ NO.4) and 2 (SEQ NO.5) The pBDA promoter DNA fragment was amplified under the conditions of denaturation at 94°C for 45 seconds, annealing at 54°C for 45 seconds, extension at 72°C for 1 minute, repeating 30 cycles, and extension at 72°C for 5 minutes. Mix the resulting pBDA DNA fragment with the secretory signal peptide nucleic acid sequence Xs, add high-fidelity DNA polymerase and dNTP, denature at 94°C for 1 minute, anneal at 60°C for 2 minutes, and repeat 8-10 cycles, then use primer 1 and Primer 3 (SEQ NO.6) was used to amplify the full-length nucleic acid fragment of the pBDA / Xs fusion gene by PCR...
example 2
[0078] The preparation of example 2 transformant oral liquid of the present invention
[0079] 1. Preparation of Seed Bacteria
[0080] 1.1 Amplification culture: take the OXM gene stably transformed bifidobacterium bacterial liquid identified by PCR detection, Gram staining and induction expression experiment, inoculate in 10ml improved MRS medium (containing 0.05% cysteine, 100ug / ml ampicillin), anaerobic culture at 37°C; when the OD695 absorbance value of the bacterial suspension reaches 0.6, centrifuge at 4500 rpm for 5 minutes to harvest the bacteria;
[0081] 1.2 Frozen storage:
[0082] 1.2.1 Freeze storage method 1: add an appropriate amount of MRS culture medium containing 15-50% glycerol or 10-20% dimethyl sulfoxide (DMSO) antifreeze, and freeze at -20 to -80°C after aliquoting as seeds bacteria.
[0083] 1.2.2 Cryopreservation method 2: add glycerin, 10% (w / v) skimmed milk powder, 3% (w / v) lactose and 3% (w / v) final concentration of glycerin Freeze-dry the freez...
example 3
[0088] Example 3 freeze-dried powder
[0089] Get the bacterium precipitation that 2.1 method harvests in the example 2, add the xylo-oligosaccharide solution resuspended bacterium that final concentration is 20% (w / v), then add the glycerol that final concentration is 15% (w / v), 10% ( w / v) of skimmed milk powder, 3% (w / v) of lactose and 3% (w / v) of trehalose, formulated to not less than 10 per gram 8 The mixture of viable bacteria is divided into packages and then freeze-dried on a freeze-dryer, sealed and stored at 4-8°C or -20°C.
PUM
| Property | Measurement | Unit |
|---|---|---|
| Resistance | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to view more - R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap