论文部分内容阅读
本研究通过设计简并引物,并且进行温度梯度PCR和二次PCR,最终筛选出适合扩增小菜蛾Plutella xylostella(L.)中肠APN3(氨肽酶N3,Aminopeptidase N3)片段的引物和温度,这为研究APN3的基因功能、APN3同APN其他同工酶之间的亲缘关系奠定了基础。通过比较电泳图谱,发现APN3在小菜蛾中肠中的表达量远远低于APN1、APN2和APN5的表达量。小菜蛾中肠APN同工酶氨基酸序列分析发现,APN3同APN1的亲缘关系最近,同APN2的亲缘关系最远。以上结果对于揭示Bt毒素作用于小菜蛾的机理以及小菜蛾对Bt毒素的抗性分子机制具有重要理论意义。
In this study, degenerate primers were designed, and temperature gradient PCR and quadratic PCR were carried out. Finally, primers and temperature suitable for amplifying APN3 (Aminopeptidase N3) fragment of midgut of Plutella xylostella (L.) This laid the foundation for studying the gene function of APN3 and the genetic relationship between APN3 and other APN isoenzymes. By comparing the electrophoresis patterns, it was found that the expression level of APN3 in the midgut of the diamondback moth was much lower than that of APN1, APN2 and APN5. The amino acid sequence analysis of APN isozyme in the midgut of Plutella xylostella showed that APN3 has the closest genetic relationship with APN1 and the furthest genetic relationship with APN2. The above results have important theoretical significance for revealing the mechanism of action of Bt toxin on Plutella xylostella and the molecular mechanism of resistance to Bt toxin in Plutella xylostella.