摘要杂拟谷盗是一种重要的世界性储粮害虫，可危害面粉、谷物籽实等，造成巨大的农业经济损失。复眼是杂拟谷盗的主要视觉器官，由许多小眼组成，对昆虫的生长、捕食、繁殖等活动起着重要的作用。视蛋白是存在于小眼内光感受细胞膜的受体，经过长期的进化和适应，杂拟谷盗的视蛋白进化为长波（LW）视蛋白和紫外线敏感（UV）视蛋白两类。为了解杂拟谷盗LW视蛋白基因的分子进化，本实验采用RACE技术克隆得到了杂拟谷盗LW视蛋白基因的cDNA序列全长，然后与其它昆虫已提交的视蛋白基因序列进行比对和同源性分析，构建相对应的系统发育树。结果显示：克隆得到杂拟谷盗LW视蛋白基因的cDNA序列全长为1319bp，包含一段1134bp的开放阅读框，其编码蛋白含有377个氨基酸；系统发育树表明杂拟谷盗与其他一些鞘翅目昆虫LW视蛋白基因同源性较高，其中杂拟谷盗与赤拟谷盗聚为一支，显示它们种间亲缘关系最近。杂拟谷盗视蛋白基因的克隆与序列分析，为深入研究视蛋白在储粮害虫趋光行为中所发挥的作用奠定基础，不仅在理论上具有重要的意义，在实际应用中也有潜在价值，可能为储粮害虫的生物绿色防治提供新的思路和方法。81143

毕业论文关键词：杂拟谷盗  复眼  LW视蛋白  RACE  序列分析 

Molecular Cloning and Sequence Analysis of LW Opsin in Tribolium confusum

Abstract Tribolium confusum is an important world stored grain pest which can do harm to flours, grains and other seeds, causing huge economic loss in agriculture。 Compound eye is the main vision organ of T。 confusum, composed of subunits called ommatidia, which plays a significant role in its growth, foraging, finding mate, etc。 Opsins are the cell-membrane receptors of photoreceptor cells in ommatidia。 After a long period of evolution and adaptation, opsins of T。 confusum can be assigned to two spectral classes: long-wavelenth (LW) opsin and ultraviolet (UV) opsin。 The full length of LW-opsin gene sequence of T。 confusum was cloned by rapid amplification of cDNA ends (RACE)。 Then in order to learn molecular evolution of LW-opsin gene, the sequence was compared with some opsin gene sequences of other insects which had submitted to GenBank and the corresponding phylogenetic tree was constructed。 The results showed that LW-opsin cDNA sequence was 1319 bp in length, containing a 1134 bp open reading frame, which encodes 377 amino acids residues。 The analysis of phylogenetic relationship revealed that LW-opsin genes of T。 confusum and T。 castaneum were highly homologous, and T。 confusum were more closely related to T。 castaneum。 The molecular cloning of LW-opsin gene and sequence analysis lay the foundation for further study on the role of opsin in phototaxis behavior of grain pests。 It not only has important significance in theory, there are also potential value in practical applications and may provide new ideas and methods for the prevention and elimination of pests in stored grain。

Key Words: Tribolium confusum  Ommateum  Long-wavelenth (LW) opsin  RACE  Sequence Analysis

目  录

摘要 Ⅰ

Abstract Ⅱ

目录 Ⅲ

引言 1

1。材料与方法 2

1。1 供试昆虫 2

1。2 杂拟谷盗头部总RNA的提取 3

1。3 cDNA第一条链的合成 3

1。4 TDT加尾反应 3

1。5 LW视蛋白基因的克隆