Spectinomycin

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[edit] Mode of Action

Spectinomycin is a bacteriostatic agent that inhibits translocation of the peptidyl tRNA from the A site to the P site.

[edit] Working Concentrations and Stock Solutions

Up to 100mg/mL stock concentration. 100ug/mL final concentration.

[edit] Resistance Gene

Mutations in rpsE, the S5 protein, prevent spectinomycin from binding.

[edit] Inhibiting protein synthesis

In addition to being used for selection, spectinomycin can also be used to inhibit protein synthesis, for example, to inhibit wild-type ribosome activity in the presence of orthogonal ribosomes. 150-500μg/ml spectinomycin is commonly used for this purpose. Papers using spectinomycin in this way include -

1. Rackham O and Chin JW. A network of orthogonal ribosome x mRNA pairs. Nat Chem Biol 2005 Aug; 1(3) 159-66. doi:10.1038/nchembio719 pmid:16408021. PubMed HubMed [Rackham]
2. Brink MF, Verbeet MP, and de Boer HA. Specialized ribosomes: highly specific translation in vivo of a single targetted mRNA species. Gene 1995 Apr 24; 156(2) 215-22. pmid:7758959. PubMed HubMed [Brink]
3. Flynn JM, Neher SB, Kim YI, Sauer RT, and Baker TA. Proteomic discovery of cellular substrates of the ClpXP protease reveals five classes of ClpX-recognition signals. Mol Cell 2003 Mar; 11(3) 671-83. pmid:12667450. PubMed HubMed [Flynn]