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β-微管蛋白在昆虫生长发育、信号传导、抗药性等方面具有重要作用。本研究以小地老虎Agrotis ypsilon(Rottemberg)3龄幼虫为材料,利用RT-PCR、cDNA末端快速扩增技术(RACE)克隆得到小地老虎β-微管蛋白基因的cDNA序列,命名为AgTubB(GenBank登录号:JN029962),并检测了该基因在不同生长发育阶段及不同器官中表达的情况。序列分析表明:小地老虎β-微管蛋白基因的cDNA序列包含1724个碱基,包括一个1347个碱基的开放阅读框,编码448个氨基酸组成的多肽,理论蛋白分子量为50.30kD,等电点为4.81。同源性分析表明,小地老虎的微管蛋白基因与棉铃虫Helicoverpa armigera微管蛋白基因的亲缘关系最近(氨基酸序列一致性高达99.55%),与本研究所比对的其他昆虫的β-微管蛋白基因也具有高度的同源性(氨基酸序列一致性90%～99%)。在小地老虎不同发育阶段和不同器官中都检测到了β-微管蛋白基因的表达。本研究对深入研究该基因功能以及筛选微管蛋白抑制剂有重要意义。
Beta-tubulin plays an important role in insect growth, signal transduction and drug resistance. In this study, the third instar larvae of Agrotis ypsilon (Rottemberg) were used as materials to obtain the cDNA sequence of the β-tubulin gene from Agrotis ypsilon (Rottemberg) by RT-PCR and rapid amplification of cDNA ends (RACE) GenBank accession number: JN029962), and detected the gene expression in different stages of growth and different organs. Sequence analysis showed that the cDNA sequence of the β-tubulin gene of Toxoplasma gondii contains 1724 bases including a 1347 base open reading frame and encodes a polypeptide of 448 amino acids with a molecular weight of 50.30 kD and an isoelectric The point is 4.81. Homology analysis showed that the microtubule genes of D. tiger were closest to the Helicoverpa armigera tubulin gene (the amino acid sequence identity was as high as 99.55%), and β-micrometastases of other insects in this study Tubulin gene also has a high degree of homology (amino acid sequence identity 90% to 99%). The expression of β-tubulin gene was detected in different developmental stages and in different organs of Tigersia. This study is of great significance to further study the function of this gene and to screen for tubulin inhibitors.