Preparation method and application of Coxsackie virus B3 virus-like particles
A technology of coxsackie virus and type 3 virus, applied in the direction of botany equipment and methods, biochemical equipment and methods, applications, etc., can solve problems such as danger, incomplete inactivation, and disease of the inoculated
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0043] Example 1, Codon optimization and total gene synthesis of CVB3 P1 gene
[0044] The codon optimization of CVB3 P1 gene and the process of whole gene synthesis are described in detail below step by step.
[0045] 1. Codon optimization of CVB3 P1 gene
[0046] According to the wild-type CVB3 P1 gene sequence (named as CVB3P1W gene, as shown in sequence 6 in the sequence listing), the optimized CVB3 P1 gene sequence (named as CVB3P1Y gene, as shown in sequence 6) suitable for expression in yeast cells was obtained after design and repeated verification. Sequence 1 in the sequence list), that is, the P1 gene sequence of the CVB3 virus is transformed into a gene sequence containing such as Sharp PM, Cowe E.Synonymous Codon Usage in Saccharomyces cerevisiae. Yeast, 1991, 7( 7):657-678.) The optimal sequence of yeast preferred codons described, thereby improving the expression levels of four structural proteins of CVB3 VP1, VP2, VP3 and VP4 in the yeast cell culture environme...
Embodiment 2
[0051] Embodiment 2, the construction of the yeast recombinant expression vector carrying CVB3 VP1, VP2, VP3 and VP4 genes
[0052] 1. Preparation of Escherichia coli DH5α competent cells
[0053] Pick a single DH5α colony (Takara company, catalog number D9057S) from a fresh plate cultured at 37°C for 16-20h, inoculate it in 5ml of LB medium without antibiotics, and culture it at 37°C with vigorous shaking overnight (12-16h). . The next day, draw 0.5ml from the above-mentioned culture and transfer it to 50ml LB medium at a volume ratio of 1:100 to continue culturing for about 3 hours until the OD600 value of the bacterial solution is 3, and transfer the bacteria to a sterile medium under aseptic conditions. , Place in a 50ml centrifuge tube pre-cooled with ice, and bathe in ice for 30 minutes. Centrifuge at 4000rpm for 10min at 4°C, discard the supernatant, invert the tube for 1min to drain the residual culture solution, and use 10ml of ice-cooled 100mM CaCl 2 The bacterial...
Embodiment 3
[0076] Example 3, Mass expression and purification of recombinant CVB3 virus-like particles in yeast cells
[0077] 1. Preparation of Competent Cells of Saccharomyces cerevisiae
[0078] Pick a monoclonal colony INVSc1 (Saccharomyces cerevisiae, catalog number: C81000, purchased from Invitrogen) from a S. Add a suitable volume of culture into 48ml YPD culture medium and mix well to make its OD600 value 0.5. After shaking and culturing at 30°C for 1 hour, the OD600 value is 0.7. After continuing to cultivate for 30 minutes, transfer the culture to a 50ml sterile centrifuge tube , centrifuge at 1500rpm at room temperature for 5min, discard the supernatant, resuspend the pellet with 10ml of solution I (included with the kit) in Sc EasyComp transformation kit (purchased from Invitrogen), centrifuge at room temperature at 1500rpm for 5min, carefully discard the supernatant, and use 1ml of solution II ( The kit comes with) to resuspend the pellet, put 50 μl / tube into a sterilized 1...
PUM
Property | Measurement | Unit |
---|---|---|
diameter | aaaaa | aaaaa |
diameter | aaaaa | aaaaa |
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