Gene encoding polymer synthase and a process for producing polymer
A technology of polymers and encoding, applied in the fields of botanical equipment and methods, biochemical equipment and methods, genetic engineering, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0059] Cloning of polymer synthase genes from chromobacteria.
[0060] First, the genomic DNA repository does not contain the pigmented bacterium sp.USM2. We put the pigment bacterium sp.USM2 in 50 ml rich nutrient medium (1% peptone, 1% meat extract, 0.5% yeast extract, PH7.0) to culture at 30 degrees, and then the chromosome can be obtained from the microorganism by standard method DNA.
[0061] We prepared a probe to obtain a polymer synthase-containing DNA fragment from the pigmented bacterium sp.USM2. Two region-specific oligonucleotides were designed using NCBI database as a reference, and SEQ ID No. 3 and No. 4 were synthesized.
[0062] We used these oligonucleotides as primers and the chromosomal DNA of pigmented bacteria as a template to amplify the polymer synthase gene by PCR. PCR was performed for 30 cycles, each cycle including reaction at 95°C for 20 seconds, 60°C for 180 seconds, and 60°C for 180 seconds.
[0063] A 1.7kbp ApaI-SalI nucleic acid sequence in...
Embodiment 2
[0065] Preparation of hookworm transformants
[0066] The ApaI and SalI polymer synthase gene fragments were written into the cloning vector pGEM-T (agarose), which was previously cut with the same restriction enzymes. This fragment was then digested by ApaI and SalI restriction endonucleases. The final product ApaI-SalI polymerase gene fragment is written into the recombinant vector pBBR1MCS-2 (this vector can be expressed in the microorganisms of the genus Cuprophorus), and the recombinant plastid is transcribed into the PHB-4 of the genus Ancylostoma ( DSM 541) (strain unable to synthesize polymer).
[0067] First, we transformed Escherichia coli S17-1 with plasmid recombinants by the calcium chloride method. From this, we obtained Escherichia coli recombinant and recombined Hookworm C. PHB-4. We first cultured the recombinant Escherichia coli and C. hookworm PHB-4 in medium containing 1.5 ml of LB and nutrient-enriched medium respectively overnight, and then mixed 1 ml ...
Embodiment 3
[0070] Polymers were synthesized using a transformant of C. nectar.
[0071] We inoculate in 50 milliliters of mineral culture medium (3.32g / L disodium hydrogen phosphate, 2.8g / L potassium dihydrogen phosphate, 0.54g / L urea) with each copper greedy hookworm H16, copper greedy hookworm transformation strain, This medium contains 1ml / L trace elements. Put all these substances into flasks and incubate at 30 degrees. We added 50mg / L kanamycin to the transformant strain of C. hookworm, and cultured these microorganisms for 48-72 hours.
[0072] We inoculated each bacterial strain of H16 species, the transformed strain of C. nectar and PHB-4 into the above-mentioned mineral medium containing 5g / L fructose and palm kernel oil, and each strain was cultured in a 250ml flask at 30°C for 72 hours.
[0073] We added sodium valerate (2.5 g / L) for 3-hydroxyvaleric acid (3HV) production. Add 50 mg / L kanamycin to the transformed strain of Copper greedy hookworm. Add different concentratio...
PUM
Abstract
Description
Claims
Application Information
- 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, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com