Tardigrade expression cloning screens

We sought to identify new protectants from tardigrades that were sufficient to improve desiccation tolerance of E. coli. We designed and implemented an expression cloning screen to clone tardigrade cDNAs into bacterial expression vectors. After inducing protein expression in pools of bacteria carrying tardigrade cDNAs, the bacteria were desiccated. We allowed for outgrowth of survivors and then isolated plasmids from controls and desiccated samples. The plasmid inserts carrying the tardigrade cDNA were amplified by low-cycle PCR. We then sequenced these PCR fragments to determine enrichment of cDNAs following desiccation.