Abstract: The performance of species resistance to Aspergillus by conventional breeding are unstable, therefore, genetic engineering is used to breed varieties resistant to Aspergillus. CHI and GLU are broad-spectrum resistance genes and have synergistic effect. Based on constructed vectors pSC1300-8-CHI and pSC1300-13-GLU, NcoⅠrestriction site was added to the front of the promotor 8# of CHI gene and AflⅡrestriction site was added to the end of T-nos sequence through PCR amplification, ApaⅠrestriction site was added to the front of the promotor 8# of GLU gene and SpeⅠrestriction site was added to the end of T-nos sequence through PCR amplification, two genes will be connected to antibiotics self-removing carrier pLoxp after digestion, antibiotic induced-removing plant bivalent expression vectors pLoxp-8-CHI-13-GLU was constructed. Identified by restriction endonuclease, the expression vector was successfully constructed.