Bacterial Genomic Integration Kits – Gram-Negative (KanR, CamR) & Gram-Positive (KmR, EmR)

Kit Overview

Our Genomic Integration Kits for bacteria enable efficient transposon mutagenesis in both Gram-negative and Gram-positive strains. Selectable markers include KanR, CamR, KmR, and EmR. Each kit comes with hyperactive v.3 MuA transposase complexes, sequencing primers, and detailed protocols.

Gram-Negative Kits	Gram-Positive Kits
Product ID: D070201 Bacterial Genomic Integration Kit (Gram-negative, KanR) – Kanamycin resistance Price: 520 € (VAT 0%) Package Size: 10 reactions Buy Now	Product ID: D070401 Bacterial Genomic Integration Kit (Gram-positive, KmR) – Kanamycin resistance Price: 520 € (VAT 0% Package Size: 10 reactions Buy Now
Product ID: D070101 Bacterial Genomic Integration Kit (Gram-negative, CamR) – Chloramphenicol resistance Price: 520 € (VAT 0%) Package Size: 10 reactions Buy Now	Product ID: D070301 Bacterial Genomic Integration Kit (Gram-positive, EmR) – Erythromycin resistance Price: 520 € (VAT 0%) Package Size: 10 reactions Buy Now

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Applications

Insert an antibiotic-resistance marker gene into bacterial genomes
Generate in vivo knock-outs efficiently
Build genome-wide single-insertion transposon mutant libraries
Map integration sites using included transposon-specific primers

Benefits

Simple: ready-to-use Mu DNA transposition complexes—just electroporate
Efficient: generate thousands of mutants from a single reaction
Versatile: works across diverse bacterial species

Proven Across Bacterial Species

Mu DNA transposition complexes have been successfully applied to numerous bacterial species, including Gram-negatives (E. coli, Salmonella enterica, Erwinia carotovora, Yersinia enterocolitica, and Pseudomonas aeruginosa) and Gram-positives (Staphylococcus aureus, Streptococcus pyogenes, Streptococcus suis, and Clostridium perfringens). Integration efficiency primarily depends on the competence of the strain.

General Workflow

Electroporate transposition complexes into your bacterial strain
Select integration clones on antibiotic selection plates
Screen mutants using phenotypic assays, PCR, or sequencing
Map insertion sites with included primers

Figure 1 (click to enlarge). Schematic of Mu DNA transposition reaction with precut mini-Mu transposon. The desired DNA sequence (i.e. selection marker, origin of replication, other DNA sequence) is flanked by 50 bp Mu Ends (R1 and R2 MuA binding sites). Target DNA can be genomic DNA or purified plasmid DNA, and transposition reaction can be accomplished in vivo or in vitro. Integration generates 5 bp target site duplication (TSD).

References

Lamberg A, Nieminen S, Qiao M, Savilahti H (2002) Efficient insertion mutagenesis strategy for bacterial genomes involving electroporation of in vitro-assembled DNA transposition complexes of bacteriophage Mu. Appl Environ Microbiol 68:705-712
Li, Y., Xia, H., Bai, F., Xu, H., Yang, L., Yao, H., Zhang, L., Zhang, X. et al. 2007. Identification of a new gene PA5017 involved in flagella-mediated motility, chemotaxis and biofilm formation in Pseudomonas aeruginosa. FEMS Microbiol Lett 272, 188–195.
Pajunen MI, Pulliainen AT, Finne J, Savilahti H (2005) Generation of transposon insertion mutant libraries for Gram-positive bacteria by electroporation of phage Mu DNA transposition complexes. Microbiology 151:1209-1218
Lanckriet A, Timbermont L, Happonen LJ, Pajunen MI, Pasmans F, Haesebrouck F, Ducatelle R, Savilahti H, Van Immerseel F (2009) Generation of single-copy transposon insertions in Clostridium perfringens by electroporation of phage Mu DNA transposition complexes. Applied and Environmental Microbiology 75:2638–2642.