论文部分内容阅读
甜菜夜蛾核型多角体病毒(Spodoptera exigua multicapsid nucleopolyhedrovirus,SeMNPV)是对甜菜夜蛾专性的杆状病毒。egt基因是杆状病毒非必需的早期基因,它编码一种叫做蜕皮甾体尿苷二磷酸葡糖转移酶(ecdysteroid UDP-glucosyl-transferase,EGT)的多肽。采用了SDS-蛋白酶K-饱和苯酚的方法进行SeMNPV基因组DNA的提取分离,并通过PCR扩增分离出egt基因,插入T载体,转化大肠杆菌,提取重组质粒,酶切鉴定,证实,从SMNPV病毒基因组中克隆到了egt基因,序列测定SeNPV egt基因orf为1572bp,可编码523个氨基酸残基。采用DNAstar软件分析,与7种核多角体病毒及1种颗粒体病毒egt基因的同源性比较显示,SeMNPV egt基因与核多角体病毒egt序列同源性较高,其中与黄地老虎核型多角体病毒(AsNPV)同源性最高,核苷酸和氨基酸序列同源性分别为75%和79%。一些与egt结构和功能密切相关的氨基酸是绝对保守的。根据egt氨基酸序列进行分子进化树分析,可将昆虫杆状病毒分成两大类,即核型多角体病毒和颗粒体病毒。该研究为杆状病毒egt基因的遗传改造奠定了理论基础。
Spodoptera exigua multicapsid nucleopolyhedrovirus (SeMNPV) is a baculovirus that is specific to beet armyworm. The egt gene is an early non-essential baculovirus that encodes a polypeptide called ecdysteroid UDP-glucosyl-transferase (EGT). SeMNPV genomic DNA was extracted and separated by SDS-protease K-saturated phenol method. The egt gene was isolated by PCR amplification, inserted into T vector, transformed into E. coli, and the recombinant plasmids were extracted and identified by restriction enzyme digestion. The egt gene was cloned into the genome. The sequence of orn was 1572 bp, encoding 523 amino acid residues. DNAstar software analysis, and seven kinds of nuclear polyhedrosis virus and a particle virus egt gene homology comparison, SeMNPV egt gene and nuclear polyhedrosis virus egt sequence homology, and yellow tiger karyotype The polyhedrosis virus (AsNPV) has the highest homology of 75% and 79% of the nucleotide and amino acid sequences, respectively. Some amino acids closely related to the structure and function of egt are absolutely conserved. According to the amino acid sequence of egt, molecular phylogenetic tree analysis can be divided into two major types of insect baculoviruses, namely, nuclear polyhedrosis virus and granulovirus. The research laid the theoretical foundation for the genetic transformation of the egt gene of baculovirus.