A mouse gene, designated prtb (proline codon-rich transcript, brain expressed) was identified and characterized from a gene trap embryonic stem cell line. It encodes a proline-rich protein of 168 amino acids that shares 99% amino acid sequence identity with its human homologue and is located on the distal region of mouse chromosome 15. To determine the expression pattern and function of prtb, mice that carry the prtb(gt) allele were generated. During embryogenesis,prtb gene expression as revealed by beta-galactosidase (beta-gal) marker gene activity was highly regulated. Between embryonic day (E) 11.5 and E12.5, beta-gal activity was restricted to the developing heart. From E13.5 on, expression in the heart was extinguished. However, very strong beta-gal activity could be detected in the brains of adult mice, suggesting a role for this gene in brain function. Mice homozygous for the mutation were viable, fertile, and did not display any obvious abnormalities. This could be due to functional redundancy as Northern blot hybridization analysis clearly demonstrated that prtb(gt) is likely to be a null allele.