Title:
WILD-TYPE-MUTANT Π PROTEIN SWITCHING EXPRESSION SYSTEM CAPABLE OF INCREASING EFFICIENCY OF PREPARING SCREENING-TAG-FREE PLASMID
Document Type and Number:
WIPO Patent Application WO/2023/241567
Kind Code:
A1
Abstract:
Provided is a precursor plasmid used for preparing a screening-tag-free plasmid, the precursor plasmid comprising: (1) a conditioned replication initiation site having a plasmid replication initiation capacity that is dependent on regulatory proteins, wherein the conditioned replication initiation site has a first replication initiation state in the presence of a first regulatory protein and a second replication initiation site in the presence of a second regulatory protein, and has a stronger ability to initiate plasmid replication in the second replication initiation state than in the first replication initiation state; 2) a first regulatory protein expression cassette expressing the first regulatory protein; 3) a sequence encoding a repressor protein; 4) a screened tag gene; 5) a target gene, or a cloning site for inserting the target gene; and 6) paired recombination sites. Also provided are a host cell suitable for the recombination of the precursor plasmid and the production of a screening-tag-free plasmid, and a method for large-scale production of the screening-tag-free plasmid.
Inventors:
ZHOU FAN (CN)
ZHANG LU (CN)
LI HONG (CN)
ZHANG LU (CN)
LI HONG (CN)
Application Number:
PCT/CN2023/099902
Publication Date:
December 21, 2023
Filing Date:
June 13, 2023
Export Citation:
Assignee:
NANJING GENSCRIPT BIOTECH CO LTD (CN)
International Classes:
C12N15/64; C12N5/10; C12N15/63
Domestic Patent References:
| WO2016105405A1 | 2016-06-30 | |||
| WO2022217934A1 | 2022-10-20 |
Foreign References:
| US20070031378A1 | 2007-02-08 | |||
| CN1717476A | 2006-01-04 | |||
| US20010014476A1 | 2001-08-16 |
Other References:
RIEDL, A. ET AL.: "Novel conditional plasmids regulated by chemical switches provide versatile tools for genetic engineering in Escherichia coli", PLASMID, vol. 111, 10 September 2020 (2020-09-10), XP086262138, DOI: 10.1016/j.plasmid.2020.102531
JECHLINGER WOLFGANG; AZIMPOUR TABRIZI CHAKAMEH AND LUBITZ WERNER: "Minicircle DNA immobilized in bacterial ghosts: in vivo production of safe non-viral DNA delivery vehicles", JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY, vol. 8, no. 4, 1 January 2004 (2004-01-01), Karger, CH, pages 222 - 231, XP009133716, ISSN: 1464-1801
JECHLINGER WOLFGANG; AZIMPOUR TABRIZI CHAKAMEH AND LUBITZ WERNER: "Minicircle DNA immobilized in bacterial ghosts: in vivo production of safe non-viral DNA delivery vehicles", JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY, vol. 8, no. 4, 1 January 2004 (2004-01-01), Karger, CH, pages 222 - 231, XP009133716, ISSN: 1464-1801
Attorney, Agent or Firm:
CHENG & PENG INTELLECTUAL PROPERTY LAW OFFICE (CN)
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