The Function Of IAP (Inhibitor Of Apoptosis) Of SeNPV And Their Host In Mammalian Cells

This thesis paper includes five chapters.Chapter One is a brief review on Spotoptera exigua,baculovirus,IAP and CB.It focuses on the prevention of Spotoptera exigua, the character and application ofbaculovirus and the function of IAP and CB.Chapter Two focuses on the function of Spodoptera exigua IAP in 293 humanembryonic kidney (293 HEK) cells. In this study, we demonstrated that 293 HEKcells undergo apoptosis during exposure to cisplatin. Viability studies were performedfor cells expressing each wild-type protein and vector on transiently transfected cellsafter induction of apoptosis by cisplatin treatment. Contrast with the expression of thevector and the initial cells, expression of IAP inhibited apoptosis significantly. Withthe aid of an N-terminal GFP fusion to the protein, the condition of transfection anddistribution within the cell was visualized.Chapter Three focuses on the function of SeNPV IAP2 and IAP3 in 293 HEKcells. The iap2 and iap3 genes of SeNPV were amplified by PCR from the virusgenome. The PCR product was cloned into pEGFP-C1 restriction enzyme sites: EcoRI/BamH I. Contrast with the viability of the vector transfected cells and the initialcells which were induced apoptosis by cisplatin treatment. We demonstrated that thecells which expressed lAP inhibited apoptosis significantly. With the aid of anN-terminal GFP fusion to the protein, the condition of transfection and distributionwithin the cell was visualized.Chapter Four focuses on the acquirement of Helicoverpa armigera iap gene.Total RNA was abstracted from the fat body of Helicoverpa armigera and reverse transcripted to obtain complete cDNA. A pair of primers was designed according tothe homologous sequences of the genes incoding IAP which belong to Trichoplusia niand Spodoptera frugiperda. After PCR a fragment was acquired and was found to bethe conserved sequence of IAP through the result of BLASTx. Baced on this result, 3'race and 5' race were performed. The fragments we gained were cloned to pMD 18-Tand are sequencing now.Chapter Five focuses on the acquirement of Spodoptera exigua cathepsin b (cb)gene. Total RNA was abstracted from the blood lymph of Spodoptera exigua andreverse transcripted to obtain complete cDNA. Primers were designed according tothe cb sequence we have obtained before. 5' race was performed. The fragments wegained were cloned to pMD18-T and are sequencing now.