Title:
MYCOBACTERIUM TUBERCULOSIS INTEIN SPLICING INHIBITOR SCREENING SYSTEM, AND CONSTRUCTION METHOD THEREFOR AND USE THEREOF
Document Type and Number:
WIPO Patent Application WO/2023/272791
Kind Code:
A1
Abstract:
Provided is a Mycobacterium tuberculosis intein splicing inhibitor screening system. By means of taking Mycobacterium smegmatis as a type strain, a gene sequence of a protein regulated and controlled by an intein is inserted into position 65S of a kanamycin resistance protein gene, and a recombinant plasmid is constructed using a vector having a first antibiotic resistance, transferred into Mycobacterium smegmatis and then cultured in a culture medium containing a first antibiotic and kanamycin to obtain the Mycobacterium tuberculosis intein splicing inhibitor screening system, wherein the first antibiotic is any antibiotic other than kanamycin. By means of inhibiting the splicing activity of intein, the activity of a kanamycin resistance protein is affected, so that the growth of recombinant Mycobacterium smegmatis in a culture medium containing kanamycin is affected, drugs having an inhibitory effect on intein are screened out, and simple, efficient and rapid screening of new anti-tuberculosis drugs is achieved.
Inventors:
QI XINGMEI (CN)
XIONG SIDONG (CN)
LI YIYANG (CN)
ZHANG JIALAN (CN)
XIONG SIDONG (CN)
LI YIYANG (CN)
ZHANG JIALAN (CN)
Application Number:
PCT/CN2021/106637
Publication Date:
January 05, 2023
Filing Date:
July 16, 2021
Export Citation:
Assignee:
UNIV SOOCHOW (CN)
International Classes:
C12N15/74; C12N15/31; C12N15/65; C12Q1/02; C12R1/34
Domestic Patent References:
| WO2005056754A2 | 2005-06-23 |
Foreign References:
| US20120270322A1 | 2012-10-25 | |||
| US20080187922A1 | 2008-08-07 | |||
| US20020164573A1 | 2002-11-07 |
Other References:
HU JING-PING, JIANG YU-TING, QI XING-MEI: "A screening system based on kanamycin resistance against protein splicing of the ＲecA intein of Mycobacteriun tuberculosis", JOURNAL OF BIOLOGY, vol. 37, no. 2, 30 April 2020 (2020-04-30), pages 101 - 103, XP093019617, ISSN: 2095-1736, DOI: 10.3969/j.issn.2095-1736.2020.02.101
WOODS DANIEL, VANGAVETI SWETA, EGBANUM IKECHUKWU, SWEENEY ALLISON M., LI ZHONG, BACOT-DAVIS VALJEAN, LESASSIER DANIELLE S., STANGE: "Conditional DnaB Protein Splicing Is Reversibly Inhibited by Zinc in Mycobacteria", MBIO, AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 11, no. 4, 25 August 2020 (2020-08-25), US , XP093019619, ISSN: 2161-2129, DOI: 10.1128/mBio.01403-20
WOODS DANIEL, VANGAVETI SWETA, EGBANUM IKECHUKWU, SWEENEY ALLISON M., LI ZHONG, BACOT-DAVIS VALJEAN, LESASSIER DANIELLE S., STANGE: "Conditional DnaB Protein Splicing Is Reversibly Inhibited by Zinc in Mycobacteria", MBIO, AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 11, no. 4, 25 August 2020 (2020-08-25), US , XP093019619, ISSN: 2161-2129, DOI: 10.1128/mBio.01403-20
Attorney, Agent or Firm:
CENTRAL SOUTH WELL INTELLECTUAL PROPERTY OFFICE (CN)
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