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The Cloning and Expression of IAP (Inhibitor of Apoptosis) of SeNPV and Their Host

Author: MeiChunLei
Tutor: ZhangZhongXin
School: Wuhan Institute of Virology
Course: Microbiology
Keywords: Spotoptera exigua SeNPV IAP2 IAP3 PCR Induced Expression RT-PCR Transfected
CLC: S476.13
Type: Master's thesis
Year: 2006
Downloads: 121
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Abstract


This thesis paper includes four chapters.Chapter One is a brief review on recent advances on baculovirology and the function of IAP. It focuses on the fundamental researches and applied applications about baculovirology and Spotoptera exigua Muti Nucleocapsid Nucleopolyhedrovirus (SeMNPV), and does the function of the IAP.Chapter Two is a brief review on the iap2 and iap3 genes of Spodoptera exigua nuclepolehedrovirus which were amplificated by PCR from the virus genome. The iap2 was 951bp and iap3 was 939bp, encoding IAP2 and IAP3 respectively. One RING domain and two BIRs domain contained in IAP2 while one RING and one BIR domain contained in IAP3.iap2 and iap3 were ligated into expression vector pET-28a, then were transformed into BL21(DE3) competent cell respectively. Selected single colony and add it to LB(kan), then induced the amplified culture with IPTG. There was a 40kDa unique band in SDS-PAGE gel, whose molecular was consistent to that of deduced IAP2 protein.Chapter Three focus on the cloning of a new inhibitor of apoptosis protein (IAP) homolog, Spodoptera exigua IAP, from the fat body of Spotoptera exigua。IAP contains two baculovirus IAP repeat domains followed by a RING domain. SfIAP has striking amino acid sequence similarity with baculoviral IAPs, CpIAP and OpIAP, suggesting that baculoviral IAPs may be host-derived genes. Total RNA was isolated from the fat body of Sdoptera exigua and reverse

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CLC: > Agricultural Sciences > Plant Protection > A variety of control methods > Biological control > The use of microbial pathogens > Insect viruses
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