Chicken prolactin (PRL) is a physiological candidate gene for egg production. The objective of the current research was to investigate the association of polymorphisms in the chicken PRL promoter region with egg production. Genotyping of 177 individuals from White Leghorn, Yangshan, Taihe Silkies, White Rock, and Nongdahe breeds for 6 single nucleotide polymorphisms (C-2402T, C-2161G, T-2101G, C-2062G, T-2054A, and G-2040A) and 1 24-bp indel (insertion-deletion) at the site of -358 of the chicken PRL gene revealed large breed differences in allelic frequencies for all but the T-2101G and T-2054A polymorphisms. An F2 population produced from Nongdahe x Taihe Silkies chickens consisted of 374 hens, which were recorded for egg production traits and genotyped for the above 7 polymorphisms. Marker-trait association analysis indicated that the 24-bp indel was associated with egg production (P < 0.01) and that H3 (C C T C T G) was the most advantageous haplotype for egg production.