大蒜E病毒外壳蛋白基因的原核表达及抗血清制备

The Prokaryotic Expression of The Coat Protein Gene of Garlic virus E and The Antiserum Preparation

设计特异性引物PCR扩增了余杭大蒜病样中的大蒜E病毒 (GarV E)的全长CP基因 ,构建原核表达载体并在大肠杆菌中过量表达 ,纯化表达产物后免疫家兔制备抗血清。Westernblot检测结果表明 ,抗血清与GarV E的CP起强的特异性反应。 7个大蒜样品中 ,内蒙古赤峰市、宁夏银川和甘肃天水等 4个样品受到GarV E的侵染。试验也证明了田间GarV E编码的CP分子量为 35kD ,不同于已报道的其他葱X病毒属成员的 2

Specific primers were designed to be used in amplification of the complete coat protein gene of Garlic virus E Yuhang isolate. The expected CP gene was then inserted into the pSBET vector for the prokaryotic expression. The aimed protein was purified and used to immune the rabbit for the antiserum preparation. Western blot analysis confirmed that the antiserum reacted strongly and specifically to the CP of GarV E. GarV E was detected in 4 garlic samples from Chifeng ,Yinchuan and Tianshui cities respectively. It also revealed that the molecular weight of GarV E CP was 35kD which was different to other allexiviruses(28kD).