甘蔗黄叶病毒运动蛋白的原核表达和抗血清制备

 甘蔗黄叶病毒(Sugarcane yellow leaf virus,ScYLV)属于黄症病毒科(Luteoviridae)、马铃薯卷叶病毒属(Polerovirus),主要由蚜虫传播,能够侵染甘蔗、玉米等多种作物,引发严重的植物病害。本研究将编码ScYLV运动蛋白(Movement protein,MP)的基因连接到pDB-His-MBP原核表达载体上,转化到大肠杆菌菌株Rosetta中,经IPTG诱导,表达分子量大小约为70 kDa的融合蛋白,将纯化后的融合蛋白制备多克隆抗血清。利用western blot检测抗血清的效价为1:128 000,灵敏度为1:256,抗血清不与马铃薯卷叶病毒属(Polerovirus)的其它病毒以及黄症病毒属(Luteovirus)病毒发生交叉反应,具有很好的特异性。本文制备的ScYLV MP抗血清可以用于ScYLV的检测,并为深入研究ScYLV与寄主的相互作用等问题提供材料基础。

Prokaryotic expression and antiserum preparation of the movement protein of Sugarcane yellow leaf virus

Sugarcane yellow leaf virus (ScYLV) belongs to the genus Polerovirus in the family Luteoviridae.It is mainly transmitted by aphid and it can infect varieties of plants such as sugarcane and maize,causing severe plant diseases.We cloned the movement protein (MP) gene of ScYLV Yunnan isolate and inserted into the expression vector pDB-His-MBP.The 70 kDa MP fusion protein was expressed in Escherichia coli Rosetta strain,purified protein was used to prepare polyclonal antiserum against to ScYLV MP.The results of western blotting showed that the titer of antiserum is 1:128000,the sensitivity of antiserum is 1:256.The antiserum could react with ScYLV MP specifically,but not with MP of other members of the genera Polerovirus and Luteovirus.Therefore,the prepared antiserum can be used to specially detect ScYLV and provide research materials for the further study on interaction between ScYLV and host plants.