Plasmid for high-precision gene editing of multi-drug-resistant gram-negative bacteria as well as construction method and application of plasmid

The invention relates to a plasmid for high-precision gene editing of multi-drug-resistant gram-negative bacteria as well as a construction method and application of the plasmid. The plasmid is based on an R6K gamma origin suicide replicon, is provided with a sucrose lethal gene sacB for reverse scr...

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Bibliographic Details
Main Authors LIU GUANGYU, GAO JIAXIN, FAN CHENGFENG, FAN MEIMEI, CHEN FENG
Format Patent
LanguageChinese
English
Published 08.03.2024
Subjects
BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROCESSES USING MICROORGANISMS
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
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Summary:The invention relates to a plasmid for high-precision gene editing of multi-drug-resistant gram-negative bacteria as well as a construction method and application of the plasmid. The plasmid is based on an R6K gamma origin suicide replicon, is provided with a sucrose lethal gene sacB for reverse screening, and is provided with a green fluorescent protein gene sfgfp, a kanamycin resistance gene kanR or a hygromycin resistance gene hygR for multi-stage screening of gene editing; and a homologous arm cloning site and a series of regulatory elements are supplemented. According to the method, homologous recombination and screening of genes such as sacB, sfgfp, kanR and the like are realized by constructing DNA fragments which are the same as upstream and downstream sequences of a target basic group or a gene to obtain a target strain, and the problems of long time consumption and low efficiency of gene editing aiming at multiple drug-resistant negative bacteria at present are solved. The resistance gene carried by
Bibliography:Application Number: CN202311577569