Generate random gene knockouts in non-E. coli bacteria for metagenomics and bacterial strain engineering.

• Generate mutants with improved genetics or function
• Identify genes involved in pathogenesis, toxicity, biofilm development
• Unravel metabolic pathways
• Identify essential genes and regulatory elements
• Insert Kan selectable marker into gDNA and rescue clones in E. coli host expressing the pir gene product
• 100’s of citations for many different applications

Create and sequence bacterial mutants faster with in vivo transposomics tools

Transposomes are used for in vivo mutagenesis for a broad range of bacteria, including Gram positive and Gram negative strains. The transposome is a complex of a engineered hyperactive Tn5 Transposase enzyme, and a DNA sequence (transposon) to be inserted. The transpon sequence in the Z-Tn5™ Tnp Transposome™ Kit consists of a R6Kγ origin of replication and a kanamycin resistance marker, which can be used for insertional mutagenesis and easy downstream rescue and sequencing of the transposon insertion site.

To generate a library of mutants from your organism, simply transform your bacterial strain of interest with the transposome, and grow colonies on selective (kanamycin) media. The colonies can be expanded and screened in almost any way you can imagine - with a phenotypic screen designed to identify a particular attribute, a genetic screen for a specific gene disruption event, or a functional screen (for loss of gene function). After "hits" are identified by the screen, the clone can be isolated and characterized further. The transposition insertion site can be sequenced directly by NGS, or by "rescue" cloning of the flanking sequences followed by Sanger sequencing outward from the transposon ends.

One of the advantages of transposon mutagenesis is that the transposon serves as a marker that can be used to clone and sequence the region of genomic DNA that has been disrupted. The presence of the conditional origin of replication (R6Kγori) enables the propagation or "rescue" of the region of genomic DNA or plasmid, into which the transposon has been inserted.

The power of this technology is only limited by your imagination!