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Pituitary Specific transcription factor 1 (PIT-1) gene is one of the POU gene family and play key roles in the regulation of muscle development which is important for body weight in chickens. This study was carried out to detect the polymorphism of PIT-1 gene and study its association with body weight in Fulani and Yoruba ecotype of Nigerian indigenous chickens. A total of 100 day-old chicks comprising of fifty chicks of Fulani and Yoruba ecotypes were used for this study. These chicks were genotyped using PCR-RFLP technique and their growth rate was also measured throughout the study. Our results showed two alleles A and B controlling genotypes AA and AB which were significantly associated with bodyweight of the chickens. Interestingly, we found out that AA genotype had the highest frequency of 0.64 and 0.76 compared with AB genotype which had a frequency of 0.36 and 0.24 in the Fulani and Yoruba ecotype respectively. Furthermore, the PIT-1 genotypes (AA and AB) were significantly associated (p < 0.05) with body weight at week 4 and week 12 in Fulani and Yoruba ecotype chicken respectively. PIT-1 gene was polymorphic in Fulani and Yoruba ecotypes of Nigeria Indigenous Chickens.