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Mode of Action

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

Working Concentrations and Stock Solutions

Up to 100mg/mL stock concentration in H2O. 100ug/mL final concentration.

Resistance Gene

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

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 | PubMed ID:16408021 | 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. DOI:10.1016/0378-1119(95)00030-a | PubMed ID:7758959 | 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. DOI:10.1016/s1097-2765(03)00060-1 | PubMed ID:12667450 | HubMed [Flynn]

All Medline abstracts: PubMed | HubMed