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A shuttle vector, pTOPO-C, which replicated in Escherichia coli and Clostridium beijerinckii was constructed. pTOPO-C vector was derived from pCR2.1TOPO (Invitrogen) which contained pUC origin, ampicillin resistance gene and kanamycin resistance gene. The ampicillin resistance gene was removed from pCR2.1TOPO and the replication origin of the megaplasmid, pSOL1, from the Clostridium acetobutylicum and erythromycin resistance gene for the selection marker in C. beijerinckii was inserted. We also developed a method to transform plasmid into C. beijerinckii by electroporation. This shuttle vector and transformation protocol can provide the basis for metabolic engineering of C. beijerinckii.