Abstract: A marker-free vector (pBINMF) derived from the binary vector pBINPLUS was generated, which only contained a multiple cloned site between left and right borders as validated by restriction analysis and sequencing. In order to identify the efficiency of pBINMF for transformation, the gus gene was cloned into the vector and was introduced into the tomato cultivar Moneymaker via Agrobacterium tumefaciens-mediated transformation. Gene-specific PCR and GUS stain assay showed that gus gene was integrated into the tomato genome and was expressed. This vector could become a reliable and efficient tool for direct generating marker-free transformants in plants, particularly useful for vegetatively propagated crops like potato and cassava.

Key words: marker-free transformation, vector construction, transgene, tomato