Background: Protamines are the nuclear proteins essential for chromatin compaction during spermatogenesis. Spermatozoa may develop the SDF if the spermatogenesis is dysregulated, which affects male infertility Objective: This study was conducted to quantify the PROTAMINE 1 gene expression in the semen specimen of infertile Iraqi male and estimate sperm DNA fragmentation. Patients and Material: 50 semen specimens were gathered from infertile males and 50 samples from healthy fertile individual as control group and subjected for semen analysis, determinations the expression of PROTAMINE 1 gene and estimation sperm DNA fragmentation. Results: There were no appreciable differences in the ejaculation volume between the studied groups.the fertile group's mean sperm concentration (m/ml)(49.58 ±2.93)was statistically significantly (p <0.05) higher than the infertile male group(27.08±15.7).The progressive motility of fertile male(57.20 ± 2.27) was significantly(P < 0.001) higher as compared to other infertile male groups(22.29 ± 1.95).The morphologically normal sperm of control males(52.20 ± 1.91) was a highly significant (P < 0.001) as compared to infertile male groups (36.30± 2.20).The fold protamine 1 gene expression of patients group(0.26) had a lower significantly than the healthy group (1).The mean SDF of infertile male group(43.05 ± 2.93) was statistically significant  (P < 0.001)than the fertile male group( 17.97 ± 9.90). Conclusion: The current study found that increased sperm with breaks DNA was linked to reduced protamine 1 gene expression and negative impacts on sperm parameters.