亨得拉病毒核蛋白基因(N)在大肠杆菌的高效表达与反应原性鉴定

从引进的含亨得拉病毒（Hendra virus，HeV）基因的克隆质粒中扩增得到该病毒核蛋白基因（N），将其克隆到pET-28a（＋）原核表达栽体，转化大肠杆菌后成功地表达了N蛋白，SDS-PAGE电泳分析结果表明目的基因在JM109（DE3）大肠杆菌菌株中得到了良好的表达，目的蛋白量占菌体总蛋白的14．6％。Western-blotting检测表明，表达的蛋白可以被兔抗亨得拉全病毒阳性血清识别，表明该蛋白具有良好的反应性。该研究结果可进一步应用于诊断试剂和单克隆抗体的制备以及流行病学调查，以防范该病在我国的流行。

Expression of Nucleotide(N) Protein of Hendra Virus in E. coli

As a zoonotic disease caused by Hendra virus(HeV),which emerged first in Australia in 1994,the Hendra disease characterized by the severe pathological damage on respiratory and central nervous system,usually associated with high morbidity and mortality rates in equine and human.In this study HeV N gene fragment was amplified by PCR from the cloning plasmid imported from Australia and engineered into plasmid pET-28a(+) for prokaryotic expression.The N protein was expressed effectively in E.coli,accounting for 14.6% of total bacteria proteins,as evidenced by SDS-PAGE and demonstrated an expected reactivity in Western-blotting with rabbit antiserum against the intact Hendra virus particles.The study represents a preliminary work towards the development of diagnostic reagents and monoclonal antibodies for detection of Hendra virus and the epidemiological investigation of the disease,in order to prevent its potential outbreak in China.