Serum response factor (SRF) is a ubiquitously expressed stimulus-dependent transcription factor that regulates gene expression by binding to serum response element in the promoter region of target genes. Recent studies in mice have shown that SRF is important for activity-dependent gene expression and synaptic plasticity in the adult brain but is dispensable for neuronal survival. Given these important functions of SRF in the CNS, it is expected to play a critical role in several aspects of learning and memory. Here we evaluated the role of SRF in conditioned reinforcement using two lines of conditional SRF mutant mice. These SRF mutant mice exhibited different spatial patterns of SRF deletion in the post-natal forebrain and notably within the hippocampus. SRF deletion was more widespread in SRF-CKCre mutants than in SRF-SynCre mutants, particularly in areas of the cortex and striatum. Mutant and wild-type mice were trained to associate one auditory cue (CS+) with reward, whereas a second cue remained relatively neutral (CS-). All mice readily acquired this discrimination, entering the food cup during CS+ but not during CS-. In a subsequent test of conditioned reinforcement, in the absence of food, wild-type control mice and SRF-SynCre mice learned to selectively perform an instrumental response that yielded CS+ presentation rather than another response that produced CS-. SRF-CKCre mutants failed to show this preferential responding for CS+. These results suggest a role for SRF in conditioned reinforcement, a manifestation of incentive learning that has been implicated in many aspects of adaptive and maladaptive behavior, such as substance abuse and eating disorders.