Title:
METHOD FOR EDITING GENOME OF BACTERIA CAPABLE OF PRODUCING BACTERIAL CELLULOSE
Document Type and Number:
WIPO Patent Application WO/2023/240607
Kind Code:
A1
Abstract:
Provided is a gene editing method for the genome of bacteria capable of producing bacterial cellulose. The method comprises the following steps: S1) determining a target gene or site to be modified in bacteria capable of producing cellulose membranes, and determining a first homologous arm sequence according to the target gene or site to be modified; S2) preparing a donor plasmid, wherein the donor plasmid comprises a replication initiation site, a screening gene and a second homologous arm sequence for allelic exchange with a genome; and S3) transferring the prepared donor plasmid into the bacteria capable of producing cellulose membranes, performing culturing, and performing screening by means of a screening method corresponding to the screening gene to obtain gene-edited recombinant bacteria of the bacteria capable of producing cellulose membranes, wherein the sequence of the donor plasmid, which is linearized, is inserted into a target gene or site to be modified in the genome of the recombinant bacteria. By means of the present invention, rapid and simple genetic modification is performed on the genome of bacteria capable of producing cellulose membranes by using a single recombination method.
Inventors:
GAO YANMEI (CN)
LI FEIXUAN (CN)
JIN FAN (CN)
LI FEIXUAN (CN)
JIN FAN (CN)
Application Number:
PCT/CN2022/099492
Publication Date:
December 21, 2023
Filing Date:
June 17, 2022
Export Citation:
Assignee:
SHENZHEN INST OF ADV TECH CAS (CN)
International Classes:
C12N15/74; C12N1/20; C12N15/63; C12R1/02
Foreign References:
| US20220081692A1 | 2022-03-17 | |||
| CN107299073A | 2017-10-27 | |||
| US20050273869A1 | 2005-12-08 |
Other References:
LIU LING-PU, YANG XUE, ZHAO XIANG-JUN, ZHANG KAI-YUE, LI WEN-CHAO, XIE YAN-YAN, JIA SHI-RU, ZHONG CHENG: "A Lambda Red and FLP/FRT-Mediated Site-Specific Recombination System in Komagataeibacter xylinus and Its Application to Enhance the Productivity of Bacterial Cellulose", ACS SYNTHETIC BIOLOGY, AMERICAN CHEMICAL SOCIETY, WASHINGTON DC ,USA, vol. 9, no. 11, 20 November 2020 (2020-11-20), Washington DC ,USA , pages 3171 - 3180, XP093118446, ISSN: 2161-5063, DOI: 10.1021/acssynbio.0c00450
ZHANG WEN ，, QI XIANG-JUN, LI YAN-JUN: "Studying on the pde-gene′s disruption of the Acetobacter xylinum", JOURNAL OF SHAANXI UNIVERSITY OF SCIENCE ＆ TECHNOLOGY(NATURAL SCIENCE EDITION). 2013(05) PAGE:121-125, vol. 31, no. 5, 25 October 2013 (2013-10-25), pages 121 - 125, XP093118460, DOI: 1000-5811(2013)05-0121-05
HALL P. E. ET AL.: "Transformation of Acetobacter xylinum with Plasmid DNA by Electroporation", PLASMID, vol. 28, 31 December 1992 (1992-12-31)
ZHANG WEN ，, QI XIANG-JUN, LI YAN-JUN: "Studying on the pde-gene′s disruption of the Acetobacter xylinum", JOURNAL OF SHAANXI UNIVERSITY OF SCIENCE ＆ TECHNOLOGY(NATURAL SCIENCE EDITION). 2013(05) PAGE:121-125, vol. 31, no. 5, 25 October 2013 (2013-10-25), pages 121 - 125, XP093118460, DOI: 1000-5811(2013)05-0121-05
HALL P. E. ET AL.: "Transformation of Acetobacter xylinum with Plasmid DNA by Electroporation", PLASMID, vol. 28, 31 December 1992 (1992-12-31)
Attorney, Agent or Firm:
BEIJING CHENGHUI LAW FIRM (CN)
Download PDF: