pGMβ1
(Plasmid #216202)

• Purpose: (Empty Backbone) Chromosomal gene manipulation (gene insertion, conversion, deletion) of dairy used Lactobacillus bulgaricus, a difficult gene to manipulate, is now possible by conjugation using this pGMB1 plasmid.
• Depositing Lab: Yasuko Sasaki
• Publication: Iwamoto et al Biosci Microbiota Food Health. 2022;41(1):20-29. doi: 10.12938/bmfh.2021-014. Epub 2021 Oct 18.

Backbone

• Vector backbone: pAMbeta1 and pGEM-Teasy
• Backbone manufacturer: pAMbeta1 from Enterococcus faecalis (gram positive) and pGEM-Teasy from E. coli
• Backbone size (bp): 30831
• Modifications to backbone: pAMbeta1 replicates in lactic acid bacteria but is difficult to genetically manipulate, so we shuttled it so that the plasmid could be constructed in E. coli.
• Vector type: Bacterial Expression ; Shuttle vector, Conjugal plasmid, theta type replication in lactic acid bacteria
• Promoter: lac promoter in pGEM-Teasy,　Promoters in pAMbeta1 are unknown.

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: As it is a shuttle vector, high copy / Ampicillin resistant in DHalpha, and low copy/ Erythromycin resistant, 45˚C growth in Lactic acid bacteria.
• Copy number: High Copy

Cloning Information

• Cloning method: Ligation Independent Cloning

Resource Information

• Supplemental Documents:
  • pAMbeat1 complete nuc. addgene.seq
• A portion of this plasmid was derived from a plasmid made by: pAMbeta1 plasmid was transferred from Prof. Clewell DB of Michigan University in 1987. pGEM-Teasy was purchased by ProMega co..

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Industry MTA

Depositor Comments

https://doi.org/10.1016/j.idairyj.2023.105827
Lactobacillus delbrueckii subsp. bulgaricus OLL1073R-1 eps gene knockout mutants reduced exopolysaccharide synthesis and immunomodulatory activities