【作者】 夏爱华；

【导师】 李卫国； 张志芳；

【作者基本信息】 山东农业大学 ， 特种经济动物饲养， 2006， 硕士

【摘要】 分子生物学的发展和杆状病毒载体表达系统(baculovirus expression vector systems,BEVS)的改进与完善,为成功表达各种具有生物活性的有用蛋白和疫苗提供了一条有效的途径。目前多种杆状病毒基因组全序列已被测定,而柞蚕杆状病毒表达系统由于多角体蛋白基因启动子效率比较高、表达量高,成本低,有其独特优点,因此我们对柞蚕杆状病毒基因组进行了初步研究。采用随机克隆方法,建立柞蚕杆状病毒的质粒基因文库,通过对插入片段进行克隆鉴定和序列分析,获得了柞蚕杆状病毒的三个新基因,并对其序列进行了分析。1在基因结构上,odv-e56基因上游具有晚期调控保守序列TTAAG,是一晚期表达基因,其推定的ORF为1125 bp,共编码374个氨基酸,基因内部有一个保守的糖基化位点、六个半胱氨酸残基和两个疏水区;ie-2和pe-38基因都是极早期表达基因,其推定的开放阅读框分别编码295和302个氨基酸,并且内部含有一个保守的锌指结构和亮氨酸拉链。2核苷酸和氨基酸同源性比较结果表明:柞蚕核型多角体病毒的odv-e56基因与黄杉毒蛾多角体病毒最高,而与松树叶蜂多角体病毒同源性最低;ie-2和pe-38基因都与黄杉毒蛾多角体病毒的同源性较高,与家蚕的同源性比较低。3在分子进化上,柞蚕杆状病毒odv-e56基因可能存在两种进化方式:一类以点突变为主,基因长度变化不明显,另一类突变以小片段的碱基增减为特征;ie-2和pe-38基因则出现大片段缺失,是研究物种分子进化比较典型的基因。在家蚕EST文库中,许多学者发现了有关细胞凋亡基因、组织分化基因、蛋白酶抑制剂、抗菌肽基因等许多已知功能EST,以及组织和时期特异表达基因,是家蚕分子生物学研究的活跃领域。本文通过用果蝇yellow基因的保守区MRJP检索家蚕EST文库,获得一系列同源性的EST,构建了家蚕yellow基因家族,并分析了其序列及组织表达分布。1通过用果蝇yellow基因的MRJP的保守区序列检索家蚕的EST文库,共检索到74条与其同源的家蚕EST序列,应用DNAstar、Clustal X程序进行分类拼接,获得七个都具有部分MRJP保守区的基因片段,组成一个基更多还原

【Abstract】 With the development of molecule biology the improvement of insect baculovirus expression vector systems (BEVS), it had become an efficiency approach to express active proteins and bacterins. The genomes of many kinds of baculovirus were already sequenced. As the Antheraea pernyi baculovirus expression vector system has many particular virtues,such as higher efficiency of promoter, higher expressions and lower cost , it is necessary for us to investigate the genome of Antheraea pernyi nucleopolyhedrovirus (ApNPV). we established the genomic library of radome plasmid. Results of Measuring and megaligning the inserted sequence, three new genes were obtained and analyzed.1 Genes structure were as follows : the odv-e56 gene which coded the occlusion-derived virus envelope fusion protein had a late gene motif TTAAG, its ORF had 1125 bp and encodes 374 amino acides, there were six conserved cysteine residues, a glycosylation site and two hydrophobic domains; pe-38 and ie-2 are immediate early regulator genes which their ORFs encoded 286 and 302 amino acides separately, they also had a conserved zinc finger and a leucine domain separately.2 By comparing the identities of nucleotide acid sequences and amino acid sequences with the other baculoviruses, we found that the ApNPV gene odv-e56 was highest homological with that of Orgyia pseudotsugata nuc1eopolyhedrovirus (OpNPV), but lowest homological with that of Neodiprion lecontei nucleopolyhedrovirus (NlNPV). ApNPV pe-38 and ie-2 gene were the highest homological with that of OpNPV,but lower than that of Bombyx mori nucleopolyhedrovirus (BmNPV).3 From the evolution of the nucleopolyhedrovirus, we concluded that the evolution of ApNPV odv-e56 had two kinds of methods: one was point mutation and the other was short of amino sequence; the conservetaive domain of pe-38 and ie-2 were little and more amino acids are shorted, so they were typical genes of investigating evolution and relationship in the更多还原