The Bee Microbiome Toolkit (BTK) is a Golden-Gate compatible toolkit of genetic parts designed for working with newly isolated, non-model bacteria. Like the yeast toolkit, it relies on type IIS enzymatic assembly of Type 1-8 parts with defined nucleotide overhangs into complete vectors. The toolkit includes broad-host-range (BHR) promoters and a BHR plasmid backbone that is replicated in diverse Gram-negative and Gram-positive hosts. BTK plasmids possess an origin of transfer (oriT), and can be delivered by conjugation or electroporation. Here we describe example protocols and use cases of the BTK. BTK parts will soon be available from Addgene. If you use the BTK parts or these protocols in your work, please cite: 1. Leonard, S. P. et al. Genetic Engineering of Bee Gut Microbiome Bacteria with a Toolkit for Modular Assembly of Broad-Host-Range Plasmids. ACS Synth. Biol. acssynbio.7b00399 (2018). doi:10.1021/acssynbio.7b00399.
Detailed description of BTK components and Parts List