Construction method of recombinant novel coronavirus spike protein receptor binding region type 9 adeno-associated virus
A coronavirus and spike protein technology, applied in virus/bacteriophage, microbial-based methods, viruses, etc., can solve problems such as death and lung infection, achieve high application value, prevent large-scale infection, and provide long-term effective immune protection Effect
Active Publication Date: 2021-04-06
NAT INST FOR FOOD & DRUG CONTROL
View PDF4 Cites 5 Cited by
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
After 2019-nCoV infects the human body, it will lead to lung infection and severe death
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View moreImage
Smart Image Click on the blue labels to locate them in the text.
Smart ImageViewing Examples
Examples
Experimental program
Comparison scheme
Effect test
Embodiment Construction
[0055] see figure 1 :
[0056] The insertion gene described in the present invention: the RBD region of the receptor binding region of the two novel coronavirus spike proteins is connected with a flexible Linker, and then the tPA secretion expression signal peptide is added in front, and two restriction sites of BamHI and SalI are inserted. pAAV-MCS expression vector. Gene sequence definition: double RBD gene linked by tPA secretion signal peptide. Final product after recombination: double RBD genes linked by recombinant adeno-associated virus type 9-tPA secretion signal peptide.
[0057] see figure 2 :
[0058] 1. No. 1 clone, transfected for 24 hours, cell lysate;
[0059] 2. No. 2 clone, transfected for 24 hours, cell lysate;
[0060] 3. No. 3 clone, transfected for 24 hours, cell lysate;
[0061] 4. No. 4 clone, transfected for 24 hours, cell lysate;
[0062] 5. Negative control, transfection 24 hours, cell lysate;
[0063] 6. EGFP control, 2 transfection for 24 ...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More PUM
| Property | Measurement | Unit |
|---|---|---|
| Titer | aaaaa | aaaaa |
Login to View More
Abstract
The invention discloses a construction method of a recombinant novel coronavirus spike protein receptor binding region type 9 adeno-associated virus. The method comprises the following steps: connecting two sections of RBD sequences in series by using a flexible Linker, adding tPA secretory expression signal peptide to the front, and inserting a pAAVMCS expression vector by using BamHI and SalI restriction enzyme cutting sites to obtain the recombinant novel coronavirus spike protein receptor binding region type 9 adeno-associated virus. The recombinant 9-type adeno-associated virus vector secreted and expressed by the recombinant RBD protein and the cell strain thereof can be used for preventing a novel coronavirus 2019-nCoV vector vaccine, meet the urgent need of preventing the novel coronavirus 2019-nCoV vaccine at present, and are expected to be popularized on a large scale on the premise of further safety experiment verification; the long-term effective immune protection is realized for human beings.
Description
technical field [0001] The present invention relates to a recombinant expression virus vector for expressing the receptor binding region (RBD) of the novel coronavirus spike protein, a recombinant type 9 adeno-associated virus and a preparation method thereof, and further relates to a receptor binding region expressing the novel coronavirus spike protein A recombinant adeno-associated virus vector for a vaccine against the novel coronavirus 2019-nCoV. Background technique [0002] From 2019 to 2020, the new coronavirus 2019-nCoV broke out globally, and the therapeutic drugs and vaccines against this new coronavirus have become the focus of research and development. 2019-nCoV is a positive-strand RNA virus belonging to the coronavirus family, which has never been reported in humans before. After 2019-nCoV infects the human body, it will cause lung infection, which can lead to death in severe cases. Current studies have shown that 2019-nCoV binds to the human cell receptor a...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More Application Information
Patent Timeline
Login to View More IPC IPC(8): C12N7/01C12N15/864C12N15/50C12N5/10A61K39/215A61P31/14C12R1/93
CPCC12N7/00C12N15/86C07K14/005A61K39/12A61P31/14C12N2750/14121C12N2750/14143C12N2770/20022C12N2770/20034A61K2039/5256Y02A50/30
Inventor 王军志饶春明秦玺李永红李山虎李响丁有学裴德宁刘兰史新昌于雷周勇郭莹韩春梅朱留强
Owner NAT INST FOR FOOD & DRUG CONTROL