Method for preparing animal cell galactosylated modification influenza HA (Hemagglutinin) glycoprotein from glycosyl engineering yeast
A hemagglutinin glycoprotein, yeast technology, applied in the direction of microorganism-based methods, chemical instruments and methods, biochemical equipment and methods, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0130] Example 1, Construction of Yeast Transformed by Glycosyl Modification Pathways
[0131] (1) Construction of glycoengineering yeast strains knocked out of phosphomannosyltransferase gene, phosphate synthase gene and β-mannosyltransferase gene
[0132] 1. Construction of yeast strains with phosphomannosyltransferase gene inactivation
[0133] Yeast strain GJK02 with phosphomannosyltransferase gene inactivation is to introduce the DNA molecule shown in Sequence 1 into Pichia pastoris GJK01, and undergo homologous recombination with the homologous sequence in the GJK01 genome to knock out phosphomannose in the yeast genome transferase gene, resulting yeast;
[0134] 1. Construction of phosphomannosyltransferase gene inactivation vector
[0135] The knockout plasmid pYES2-pno1 for knocking out the mannosyltransferase (PNO1) gene is to insert the knockout mannosyltransferase (PNO1) gene fragment shown in Sequence 1 in the sequence listing into the KpnI and XbaI restriction ...
Embodiment 2
[0264] Example 2. Preparation of Influenza Virus Hemagglutinin Glycoprotein or Its Polymer Nanoparticles with Gal2GlcNAc2Man3GlcNAC2 Glycosyl Structure Modification
[0265] 1. H7N9 influenza hemagglutinin glycoprotein polymer nanoparticles prepared by engineered yeast
[0266] (1), construction of recombinant expression vector
[0267] 1. According to the full-length amino acid sequence of influenza HA from Genbank (KC853766) (A / Hongzhou / 1 / 2013 (H7N9), the coding gene was optimized according to the principles of yeast preferred codons and high gene expression.
[0268] 2. Design and synthesize the following primers:
[0269] HA7-3: 5'-ATC GCGGCCGC The sequence underlined in TTAAATACAGATAGTACATCTCAT-3' is the recognition site for NotI digestion.
[0270] HA7-5: 5'-ATC TTCGAA The sequence underlined ACGATGAACACCCAAATACTGGTTTTC-3' is the NspV restriction enzyme recognition site.
[0271] 3. Using the optimized coding gene as a template, and using HA7-3 and HA7-5 as primer...
Embodiment 3
[0411] Example 3. Preparation of H7N9 influenza virus hemagglutinin glycoprotein polymer nanoparticles with GlcNAC2Man3GlcNAC2 glycosyl structure modification
[0412] 1. H7N9 influenza hemagglutinin glycoprotein polymer nanoparticles prepared by engineered yeast
[0413] (1), the construction of the recombinant expression vector: the same as that of Example 2;
[0414] (2), construction and screening of recombinant yeast:
[0415] The recombinant vector pPICZα-HA7 was introduced into the Pichia engineering strain GJK07 constructed in (4) of Example 1 by the method of (2) of Example 2 to obtain the recombinant strain GJK07 / / pPICZα-HA7.
[0416] Screening and identification methods are the same as in Example 2 (two).
[0417] (3) Engineering yeast fermentation
[0418] The recombinant bacterium GJK07 / pPICZα-HA7 was fermented according to the method in (3) of Example 2 to obtain a homogenate.
[0419] (4) Purification and identification of influenza hemagglutinin glycoprotei...
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