Title:
METHOD FOR CONSTRUCTING EUKARYOTIC EXPRESSION VECTOR OF HUMAN PAK1 PROTEIN, EXPRESSION METHOD, AND PURIFICATION METHOD
Document Type and Number:
WIPO Patent Application WO/2023/097475
Kind Code:
A1
Abstract:
A method for constructing an eukaryotic expression vector of a human PAK1 protein, comprising the steps of: 1) carrying out PCR amplification to obtain a PAK1 gene fragment; 2) linking to an eukaryotic expression vector to obtain a recombinant plasmid; and 3) carrying out transfection, transformation, and replication to obtain an eukaryotic expression vector of a human PAK1 protein. Also disclosed is a method for expressing a recombinant human PAK1 protein, further comprising step 4): transfecting a plasmid of the eukaryotic expression vector of the human PAK1 protein obtained in step 3) into eukaryotic cells for expression. Further disclosed is a method for obtaining a recombinant human PAK1 protein, further comprising step 5): culturing the positive clone obtained in the step 4) to enlarge a system, collecting cultured cells, carrying out centrifugation to collect supernatant, and carrying out purification by means of a chromatographic column to obtain a recombinant human PAK1 protein.
Inventors:
ZHANG HAOYANG (CN)
SHI JIAN (CN)
XU AIHUA (CN)
ZHU GUOFANG (CN)
SHI JIAN (CN)
XU AIHUA (CN)
ZHU GUOFANG (CN)
Application Number:
PCT/CN2021/134500
Publication Date:
June 08, 2023
Filing Date:
November 30, 2021
Export Citation:
Assignee:
SHAOXING SHOUREN MEDICAL HEALTH & TECH CO LTD (CN)
International Classes:
C12N9/12; C07K1/22; C12N5/10; C12N15/54; C12N15/66; C12N15/85
Domestic Patent References:
| WO2014150790A1 | 2014-09-25 |
Foreign References:
| CN111808833A | 2020-10-23 | |||
| CN1826130A | 2006-08-30 | |||
| CN101778641A | 2010-07-14 | |||
| US7005285B1 | 2006-02-28 |
Other References:
WU, JINBAO ET AL.: "Construction of a Eukaryotic Green Fluorescent Protein Expression Vector Carrying the PAK-1 Gene (pEGFP-C1/PAK1) and its Expression in SW480 Cells", WORLD CHINESE JOURNAL OF DIGESTOLOGY, SHIJE WEI-CHANGBINGXUE ZAZHISHE , TAIYUAN, CN, vol. 19, no. 26, 18 September 2011 (2011-09-18), CN , pages 2730 - 2734, XP009546743, ISSN: 1009-3079, DOI: 10.11569/wcjd.v19.i26.2730
DATABASE Nucleotide 21 November 2020 (2020-11-21), ANONYMOUS : "PREDICTED: Homo sapiens p21 (RAC1) activated kinase 1 (PAK1), transcri -Nucleotide -NCBI", XP093069655, retrieved from NCBI Database accession no. XM_024448560
DATABASE Nucleotide 21 November 2020 (2020-11-21), ANONYMOUS : "PREDICTED: Homo sapiens p21 (RAC1) activated kinase 1 (PAK1), transcri -Nucleotide -NCBI", XP093069655, retrieved from NCBI Database accession no. XM_024448560
Attorney, Agent or Firm:
SHENZHEN ZHONGYI UNION INTELLECTUAL PROPERTY AGENCY CO.,LTD. (CN)
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