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Cotton bollworm Helicoverpa armigera (Hübner) is a major agricultural pests worldwide, and are widely distributed. It not only cause serious harm cotton, also feeding on corn, sorghum, soybeans and other crops. Arginine kinase (arginine kinase, AK) as the key enzyme of energy metabolism in vertebrates, studies have shown that the enzyme also played an important role in the process to adapt to adverse environmental factors, immune and defense pathogen invasion. Sensitive to Bt cotton bollworm strains (96S) for materials, cloned the arginine kinase gene promoter sequences, using RNAi technology to study the role of AK in the process of the development of the cotton bollworm. Screening room with Cry1Ac 75th generation 2971-fold resistance to bollworm strain (Bt-R) were used to study the AK in vivo expression of two different strains of H. armigera resistance-related gene and physiological parameters were measured to analyze the differences that exist between them. The main findings of this paper are as follows: 1, the use of homologous sequences information than the AK gene cloned 96S strains found gene contains an internal two exons and two introns, prokaryotic expression vector and optimization of expression conditions, to obtain the a single soluble objective protein; 2, using the Genome Walking, cloned 96S strain AK gene promoter upstream to the 5 'untranslated region extending approximately 2Kb, and the biological function prediction; 3, using the Real-Time PCR, 96S strains AK gene expression analysis, we found that the expression site, and there is time and space specific expression in the highest expression of the energy demand in the intestinal parts. AK gene expression in bollworm larvae increased with the growth of the age; while entering the adult stage, the AK gene expression decreased significantly; 4, the use of two methods of feeding and injection, successful interference 96S-bollworm AK gene normal expression. Compared with the normal group, the the the interference group bollworm growth and development was inhibited, the phenomenon of developmental delay. This phenomenon is consistent with the physiological parameters were measured and ecology statistical results; using RT-PCR and Real-Time PCR analysis of the bollworm strain of Bt-R AK gene expression, and found that it is the expression of both temporal and spatial specificity expression and increase with the growth of the age. Compared with 96S cotton bollworm strains in different ages and organization, AK expression occurred significantly change; 6, physiological parameters were measured and found: compared with 96S strains 96S and two bollworm strains of Bt-R strains of Bt-R, soluble protein content in the hemolymph increased significantly, indicating that the resistance capacity of the body has been significantly enhanced; arginine kinase and alkaline phosphatase activity was significantly decreased. The intestines of the enzyme activity showed a higher activity compared with the enzyme activity in the same strain muscle. AK gene and its expression and AK protein expression at the molecular level, the experimental study, and physiological aspects of the cotton was exploring. This is conducive to the study of the the bollworm AK molecular level, and provides a theoretical basis for the prevention and treatment of H. armigera.
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