Prolactin gene polymorphism at exon 5 of prolactin receptor gene (PRLR5) was studied in Aseel birds along with its interaction with laying performance. Laying performances were recorded as age at first laying (AFE), Body Weight at First Egg (WFE), Mean Egg Weight (MEW) and Total No. of Eggs at 90 days of laying (TEN). DNA was isolated from 2-3ml not clear of Blood from 40 birds collected from wing vein. PRLR5 locus was amplified by PCR and the product was restriction digested with Bam HI and resolved on 2% Agarose gels for genotyping. The AFE (d), WFE (Kg), MEW (g) and TEN of Aseel birds in the present study were found to be 188.40±0.22, 1.57±0.03, 43.61±0.36 & 35.48±0.40 respectively. The prolactin gene locus PRLR5 showed two alleles A & B. The frequencies of A and B alleles at this locus were 0.60 and 0.40 respectively. There was no interaction of various laying performances found with alleles of PRLR5.