Comparison On Antioxidase Isozyme Of Apis Cerana Fabricius And Apis Mellifera And Molecular Cloning Of Apis Mellifera Actin Gene Promoter

Apis cerana Fabricius and Apis mellifera have many different characteristics, such as life span and behavior and disease resistance and foraging et.al. Studying enzymes, especially antioxidases isozyme and activities, their differences and changes among different developmental stages of Apis cerana Fabricius and Apis mellifera and between them have great roles for understanding protective system of enzyme and defensive function, and for studying different characteristics of Apis cerana Fabricius and Apis mellifera. The topic studied the isozymes and activites of SOD,POD,CAT and compared and analyzed them among and between different developmental stages of Apis cerana Fabricius and Apis mellifera. At the same time, to explore the effects of expression of antioxidase genes. Cloning the promoter that is suitable for need of transgene. Which is promoter of actin gene of Apis mellifera. Making preparation to construct the transgenic vector and transfer antioxidase gene to body of honeybee. The results maked clear that:1. Antioxidases have expressed at different developmental stages of Apis cerana Fabricius and Apis mellifera in the same nutritional and environmental condition. These manifested that honeybee has a protective system of enzyme to clear the free radical in the process of growth and life;2. The SOD,POD,CAT isozymes are most similar between Apis cerana Fabricius and Apis mellifera. There are more differences on SOD,POD,CAT isozyme among different developmental stages of Apis cerana Fabricius and Apis mellifera;3. The activities of SOD isozyme between Apis cerana Fabricius and Apis mellifera, larva and imago are significant differences(P<0.01),pupa is not significant(P>0.05). The activities of POD isozyme between Apis cerana Fabricius and Apis mellifera, larva is significant differences(P<0.01),pupa and imago are not significant(P>0.05). The activities of CAT isozyme between Apis cerana Fabricius and Apis mellifera, larva and pupa are not significant(P>0.05), imago of Apis mellifera is higher than Apis cerana Fabricius(P<0.05).4. Among different developmental stages of Apis cerana Fabricius and Apis mellifera, the SOD isozyme activities are significant difference(sP<0.01). The activities of SOD of larva and imago and pupa are lower and lower in Apis cerana Fabricius and Apis mellifera ; The POD isozyme activities are different(P<0.05), The activities of POD of pupa and imago and arva are lower and lower in Apis cerana Fabricius and Apis mellifera ;The CAT isozyme activities are not significant among different developmental stages of Apis cerana Fabricius and Apis mellifera(P>0.05).5. Although POD and CAT had activities, the activities of SOD setted down gradually with the days increased in the stage of imago, activities were not been detected at the last time. Which reflected that power of antioxygen of SOD in the last time of honeybee life weakened.6. Using of knowledge bioinformation and resources of bioinformatics and ditermining the sequences of actin gene of Apis mellifera,then selecting the sequences of actin gene promoter. And designing the primer and cloning successsly the sequences of actin gene from Apis mellifera lingustica Spinola genome DNA .The studying indicated that SOD,POD,CAT isozymes have more differences among Apis cerana Fabricius and Apis mellifera, which are the important factor to cause differences of life span and behavior and secreting honey and disease resistance and foraging et al between Apis cerana Fabricius and Apis mellifera. Cloning successsly the sequences of actin gene from Apis mellifera genomic DNA. Which make preparation for constructing the transgenic vector and studying gene of antioxidase in the life of honeybee by transgenic ways.