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This study aimed at transforming PUSA-RUBY, a cultivar of tomato with a new promoter trapping vector pNU435 using Agrobacterium-mediated transformation. The Agrobacterium strain LBA4404 carrying pNU435 when co-cultivated with tomato leaves, sixteen plantlets (T1) were obtained of which 6 were found to be PCR positive and among them two PCR positive plants (PT4 and PT5) upon RB TAIL-PCR, produced amplicon of ~400bp which were cloned into pTZ57R/T by T/A cloning and sequence of the RB TAIL-PCR product with M13 F/R primers upon BLAST search showed homology to right border of T-DNA. This study demands the need for generating and testing large number of independent transgenic events for promoter trapping. At the same time, these lines can be tested for promoter trapping under special conditions such as biotic and abiotic stresses.