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1. CN111793651 - Slow virus expression vector for screening COVID-19 drug and construction method thereof

Office China
Application Number 202010700324.1
Application Date 17.07.2020
Publication Number 111793651
Publication Date 20.10.2020
Publication Kind A
IPC
C12N 15/867
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
85for animal cells
86Viral vectors
867Retroviral vectors
C12N 15/65
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
65using markers
C12N 15/50
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
31Genes encoding microbial proteins, e.g. enterotoxins
33Genes encoding viral proteins
40Proteins from RNA viruses, e.g. flaviviruses
50Coronaviridae, e.g. infectious bronchitis virus, transmissible gastroenteritis virus
C12N 5/10
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5Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
10Cells modified by introduction of foreign genetic material, e.g. virus-transformed cells
CPC
C07K 14/005
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07ORGANIC CHEMISTRY
KPEPTIDES
14Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
005from viruses
C07K 2319/21
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07ORGANIC CHEMISTRY
KPEPTIDES
2319Fusion polypeptide
20containing a tag with affinity for a non-protein ligand
21containing a His-tag
C12N 5/0696
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5Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor;
06Animal cells or tissues; Human cells or tissues
0602Vertebrate cells
0696Artificially induced pluripotent stem cells, e.g. iPS
C12N 15/65
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15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
65using markers
C12N 15/86
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15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
85for animal cells
86Viral vectors
C12N 2510/00
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2510Genetically modified cells
Applicants GUANGDONG YUANXIN REGENERATIVE MEDICINE CO., LTD.
广东源心再生医学有限公司
Inventors LIN BIN
林彬
LIN ZEBIN
林泽斌
LIN XIANMING
林先明
WU ZHIMING
吴志明
WANG PING
王萍
KONG WEIWEI
孔维维
MAI JINLIAN
麦锦连
ZHOU LISHI
周丽诗
Agents 佛山帮专知识产权代理事务所(普通合伙) 44387
Title
(EN) Slow virus expression vector for screening COVID-19 drug and construction method thereof
(ZH) 用于COVID-19药物筛选的慢病毒表达载体及其构建方法
Abstract
(EN)
The invention discloses a construction method of a slow virus expression vector for screening a COVID-19 drug. The construction method comprises the following steps: taking pCW-Cas9-Blast as a framework vector, subcloning nCoVN cDNA into the framework vector, and constructing the slow virus expression vector pCW-Puro-T2A-nCoVN of novel coronavirus nucleocapsid protein, wherein the nCoVN cDNA carries a 6*his label and a puromycin resistance gene. The slow virus expression vector for screening the COVID-19 drug, constructed by the construction method, has low requirements on an experimental environment, can be operated in a conventional laboratory, can be operated at high throughput, and can be constructed and obtained in large quantity within only 2 days.

(ZH)
本发明公开了一种用于COVID‑19药物筛选的慢病毒表达载体的构建方法,其是以pCW‑Cas9‑Blast为骨架载体,将nCoVN cDNA亚克隆到所述骨架载体中,构成新型冠状病毒核衣壳蛋白的慢病毒表达载体pCW‑Puro‑T2A‑nCoVN;其中,所述nCoVN cDNA带有一个6×his标签和嘌呤霉素抗性基因。本发明所构建的用于COVID‑19药物筛选的慢病毒表达载体对实验环境要求低,常规实验室即可操作,可高通量操作,仅需2天即可构建载体并大量获得。