The DNA genome of Hyphantria cunea nuclear polyhedrosis virus was analyzed with restriction enzymes and then localization of the polyhedrin gene on the Hind III EcoRI, SmaI and BamHI DNA fragments were carried out by Southern blotting. The polyhedrin was found on the fragments of the Hind III-W and F, EcoRI-H, SmaI-A, amd BamHI-F. The Hind III and EcoRI digestion fragments of the HcNPV genome were 24, respectively. Twelve EcoRI fragments were cloned in pUC 8 and in pBR322 vectors. The polyhedrin gene was cloned in pBR322 EcoRI site which was conformed with the Southern blotting.