象耳豆根结线虫Me-mapk1基因的克隆及RNAi分析

发布时间：2018-07-04 06:42

本文选题：象耳豆根结线虫 + 促分裂原活化蛋白激酶　；参考：《热带作物学报》2016年10期

【摘要】：促分裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)是线虫体内信号转导的重要组分,在生长发育过程中具有重要作用。本研究从象耳豆根结线虫中克隆了1个促分裂原活化蛋白激酶基因Me-mapk1c DNA序列,该序列全长1 369 bp,包含1个1 185 bp的完整开放阅读框,推导编码394个氨基酸,蛋白质分子量为45.39 ku。同源性搜索比对结果发现,该基因编码的蛋白序列与南方根结线虫MI-MAPK1具有99%的一致性。通过构建原核表达载体p ET-32a-MAPK,在IPTG的诱导下得到70 ku左右的特异融合蛋白(包括大小约26 ku的标签序列)。利用RNAi技术对象耳豆根结线虫二龄幼虫Me-mapk1基因进行沉默,线虫诱导番茄根结形成的数量显著减少,推测Me-mapk1基因的下调表达有可能影响象耳豆根结线虫二龄幼虫(J2)的生长发育。
[Abstract]:Mitogen-activated protein kinase (MAPK), an important component of signal transduction in nematodes, plays an important role in the growth and development of the nematode. This study has cloned 1 mitogen activated protein kinase gene Me-mapk1c DNA sequences from the Elephant Ear bean root knot nematode, which has a full length of 1369 BP, including 1 1185. The complete open reading frame of BP, which encodes 394 amino acids, and the protein molecular weight of 45.39 ku. homologous search results, found that the protein sequence of the gene is consistent with the 99% of the southern root knot nematode MI-MAPK1. By constructing the prokaryotic expression vector p ET-32a-MAPK, the specific fusion egg of about 70 Ku is obtained under the induction of IPTG. White (including the size of the label sequence of about 26 KU). The RNAi technique was used to silence the Me-mapk1 gene of the two instar larvae of the root knot nematode. The number of root knot formation of the nematode induced tomato root formation was significantly reduced. It is speculated that the down regulation of the Me-mapk1 gene may affect the growth and development of the second instar larvae of the Elephant Ear bean root knot nematode (J2).
【作者单位】：海南大学环境与植物保护学院;海南省农业科学院植物保护研究所海南省植物病虫害防控重点实验室;中国热带农业科学院环境与植物保护研究所;
【基金】：973计划前期研究专项(No.2014CB160307) 国家自然科学基金项目(No.31360432) 海南省植物病虫害防控重点实验室开放课题
【分类号】：S432.45

【相似文献】