棉铃虫丝氨酸蛋白酶抑制剂基因的克隆及表达分析

发布时间：2018-06-23 01:46

本文选题：棉铃虫 + 丝氨酸蛋白酶抑制剂基因　；参考：《基因组学与应用生物学》2017年04期

【摘要】：本研究对棉铃虫丝氨酸蛋白酶抑制剂(Helicoverpa armigera serine protease inhibitor,Haspi)基因的基本特征、分布情况及功能进行了初步探究。通过与已报道的家蚕丝氨酸蛋白酶抑制剂基因同源比对,利用RT-PCR技术成功克隆到3条与家蚕丝氨酸蛋白酶抑制剂基因相似性高的Haspi基因,分别命名为:Haspi-5、Haspi-6和Haspi-10。实时定量PCR结果表明3个基因在棉铃虫各组织部位均有转录,但在不同组织部位处的表达水平差异显著。Haspi-5基因在表皮中的表达水平最高;Haspi-6基因在头部的转录水平远远高于其他组织部位;Haspi-10基因在前肠中表达水平最高,中肠的转录水平也相比其他组织较高。构建表达Haspi-5、Haspi-6基因的ds RNA的载体,用表达ds RNA的HT115菌液喂食,其中Haspi-5基因在棉铃虫的转录水平明显下降,约降低了6.5倍,而Haspi-6基因转录水平略有下降但并不明显,约降低了1.2倍,但是没有观察到明显的表型变化。上述研究为Haspi基因的功能及棉铃虫防治研究提供科学依据。
[Abstract]:In this study, the basic characteristics, distribution and function of Helicoverpa armigera serine protease inhibitor (Haspi) gene were preliminarily explored. By homologous comparison with the reported homologous sericulate protease inhibitor gene, 3 sericulture eggs were successfully cloned by RT-PCR technology. The Haspi gene with high similarity of white enzyme inhibitor gene was named as: Haspi-5, Haspi-6 and Haspi-10. real-time quantitative PCR results showed that 3 genes were transcribed in the tissues of Helicoverpa armigera, but the expression level of the.Haspi-5 gene was the highest in the epidermis of the.Haspi-5 gene at different tissues, and the Haspi-6 gene was transferred to the head. The level of the Haspi-10 gene was highest in the foregut and the highest in the foregut, and the transcriptional level of the midgut was higher than that of other tissues. The carrier of the DS RNA expressing the Haspi-5 and Haspi-6 gene was fed with the HT115 bacteria expressing the DS RNA, and the Haspi-5 gene in the transcriptional level of the Helicoverpa armigera was obviously reduced by about 6.5 times. The transcriptional level of Haspi-6 gene declined slightly but was not obvious, which was about 1.2 times lower, but no obvious phenotypic changes were observed. The study provided a scientific basis for the function of Haspi gene and the prevention and control of Helicoverpa armigera.
【作者单位】：河北大学生命科学学院;
【基金】：河北省教育厅重点项目(ZD2016141) 河北省自然科学基金(C2014201056)共同资助
【分类号】：S433

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