Shuttle plasmid, construction method and application

• Main Authors: WU XINYAN, GENG BINAN, YANG SHIHUI
• Format: Patent
• Language: Chinese; English
• Published: 28.02.2023
• 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

The invention relates to the technical field of plasmids, in particular to a shuttle plasmid, a construction method and application. According to the shuttle plasmid, pEZ15Asp is used as a basic plasmid, the basic plasmid is modified, and after a replicon of the basic plasmid is replaced, the shuttle plasmid has a nucleotide sequence shown as any one of SEQ ID NO.26 to SEQ ID NO.29. The shuttle plasmid can shuttle in Zymomonas mobilis and Escherichia coli, a first replication initiation region has a nucleotide sequence as shown in SEQ ID NO.1, and a second replication initiation region has a nucleotide sequence as shown in any one of SEQ ID NO.3 to SEQ ID NO.8. After 30 generations of stable passage, the growth rate of the Zymomonas mobilis transformant obtained through electric transfer is not obviously reduced, and the Zymomonas mobilis transformant can be compatible with endogenous plasmids for a long time, so that the passage stability of the shuttle plasmids is high, and the shuttle plasmids are very sui