Prepn of metallothionein
A technology of metallothionein and fusion protein, which is applied in the field of preparation of metallothionein, can solve problems such as imperfect expression conditions and purification conditions, low expression yield, and unreasonableness, and achieve low cost, good compatibility, and improved product quality. The effect of purity
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Examples
Embodiment Construction
[0013] Escherichia coli BL21 lacking some proteolytic enzymes was used as fermentation engineering bacteria.
[0014] On metallothionein and its mutant genes, two endonuclease sites, BamHI (5' end) and EcoRI (3' end), were inserted by PCR method, and then assembled on the pGEX-4T-2 plasmid, Constructed into expression plasmids.
[0015] The expression plasmid was transferred into BL21, after small-scale culture, mixed with glycerol solution (65% (V / V) glycerol, 0.1M MgSO 4 , 25mM Tris-HCl, pH 8.0) 1:1 mixed evenly to make glycerol bacteria, stored at -70°C for a long time. Before fermentation, take out the BL21 containing the expression plasmid from -70°C, insert it into fresh 2YT (containing 100 μg / mL ampicillin), culture at 37°C with rapid shaking (270-330rpm) for 7-16 hours, and transfer at a ratio of 1:100 After that, continue to expand the culture at 30~35℃ to A 600 =0.6-1.0, add IPTG to the final concentration of 0.01-1.0mM, continue to cultivate, and then add metal i...
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