Products/Services Used | Details | Operation |
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Gene Synthesis> | For native extein splicing assays with Tk-RadA-inteinΔ286–585 , this construct was commercially synthesized and subcloned into the pET45b(+) vector backbone in-frame with an N-terminal His tag (GenScript, USA). | Get A Quote |
Mutagenesis Services> | Get A Quote |
Inteins (intervening proteins), mobile genetic elements removed through protein splicing, often interrupt proteins required for DNA replication, recombination, and repair. An abundance of in vitro evidence implies that inteins may act as regulatory elements, whereby reduced splicing inhibits production of the mature protein lacking the intein, but in vivo evidence of regulatory intein excision in the native host is absent. The model archaeon Thermococcus kodakarensis encodes 15 inteins, and we establish the impacts of intein splicing inhibition on host physiology and replication in vivo. We report that a decrease in intein splicing efficiency of the recombinase RadA, a Rad51/RecA homolog, has widespread physiol... More