著名国际病毒学期刊Journal of Virology（84:12075）发表了中科院广州生物医药与健康研究院的最新研究成果。该成果首次报告成功研制了稳定携带报告基因的复制型甲型流感病毒。
近年来，流感病毒的流行以及快速变异对人类健康提出了严峻的挑战，因此流感病毒的致病机理亟待进一步阐明，人类也迫切需要更为安全有效的新型流感疫苗及抗流感药物。广州生物医药与健康研究院呼吸疾病国家重点实验室陈凌博士研究团队，通过对实验室用非致病株PR8流感病毒基因组NA片段进行改造，整合入增强型绿色荧光蛋白报告基因（EGFP），同时保留了完整的流感病毒基因组的复制型甲型流感病毒，克服了之前其他的报道均是复制缺陷型流感病毒所存在的问题。携带报告基因的复制型甲型流感病毒可以在细胞以及鸡胚中稳定生长、传代并表达报告基因，因此可以很好的的模拟野生型病毒；同时，报告基因的检测便捷可靠，可用于筛选与评价抗流感病毒药物和抗体。
更有意义的是，检测报告基因的表达情况可以用来实时追踪流感病毒在动物模型体内的感染、生长与分布。因此，该成果不但为流感病毒的基础研究，也为流感疫苗、抗流感病毒药物和抗体的研发提供了强有力的工具。
此项研究得到了中国科学院、国家自然科学基金、呼吸疾病国家重点实验室、广东省自然科学基金等资助。（生物谷Bioon.com）
生物谷推荐原文出处：
Journal of Virology doi:10.1128/JVI.00046-10
Generation of Replication-Competent Recombinant Influenza A Viruses Carrying a Reporter Gene Harbored in the Neuraminidase Segment
Feng Li,1, Liqiang Feng,1, Weiqi Pan,1 Zhenyuan Dong,1 Chufang Li,1 Caijun Sun,1 and Ling Chen1,2*
State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Biomedicine and Health (GIBH), Chinese Academy of Sciences, Guangzhou 510530,1 Department of Medical Biotechnology, University of Science & Technology of China, Hefei 230026, China2
Replication-competent influenza viruses carrying reporter genes are of great use for basic research, screening of antiviral drugs, and neutralizing of antibodies. In this study, two recombinant influenza A viruses with a neuraminidase (NA) segment harboring enhanced green fluorescent protein (EGFP) in the background of A/PR/8/34 (PR8) were generated. The viral RNA (vRNA)-specific packaging signals for NA were largely retained for efficient packaging. An "autocleave" 2A peptide sequence, which was inserted at the N terminus or the COOH terminus of NA to link with EGFP, enabled NA and EGFP to be expressed monocistronically. Further analysis demonstrated that both viruses, named rPR8-EGFP+NA and rPR8-NA+EGPF, although with some characteristic differences in growth and EGFP expression, could replicate in noncomplementary cells and propagate to large quantities while maintaining genome stability after multiple passages in embryonated eggs. These replication-competent influenza viruses carrying reporter genes are a great addition to the tool set for developing antiviral therapeutics and vaccines and for in vivo studies of viral dissemination and pathogenicity.
