The objective of this investigation was molecular population study on Penaeus semisulcatus stocks from the Persian Gulf and Oman Sea. Samples were collected using trawling method from Hormuz (40 individuals) and Bushehr (35 individuals) regions. The DNA of samples were extracted using phenol and chloroform method and then were simplified using a pair premier of Cytochrom Oxidase Subunit I (COI) gene sequence by a thermal cycler. Nine restriction enzyme were Used to digest the larger gene region that five of them (Alu I, Hinf I, Hinc I I, Hpa I I and Rca I) appeared Polymorphic patterns. Reap software and X^2 test were used to analyses the RFLP data. The average nucleotide diversity arid haplotype diversity among the population were 0.0345720 ± 0.0011952 and 0.28590±0.08174 and nucleotide divergence among population, being studied, is supposed to be 8.5%. Considering the result dispersion of haplotypes in two region showed a significant difference and this is an evidence for proving the variety of the stocks