著名國際病毒學期刊Journal of Virology（84:12075）發(fā)表了中科院廣州生物醫(yī)藥與健康研究院的最新研究成果,。該成果首次報告成功研制了穩(wěn)定攜帶報告基因的復制型甲型流感病毒,。
近年來，流感病毒的流行以及快速變異對人類健康提出了嚴峻的挑戰(zhàn),，因此流感病毒的致病機理亟待進一步闡明,，人類也迫切需要更為安全有效的新型流感疫苗及抗流感藥物。廣州生物醫(yī)藥與健康研究院呼吸疾病國家重點實驗室陳凌博士研究團隊,，通過對實驗室用非致病株PR8流感病毒基因組NA片段進行改造,，整合入增強型綠色熒光蛋白報告基因（EGFP）,，同時保留了完整的流感病毒基因組的復制型甲型流感病毒,，克服了之前其他的報道均是復制缺陷型流感病毒所存在的問題,。攜帶報告基因的復制型甲型流感病毒可以在細胞以及雞胚中穩(wěn)定生長,、傳代并表達報告基因，因此可以很好的的模擬野生型病毒,；同時,，報告基因的檢測便捷可靠,，可用于篩選與評價抗流感病毒藥物和抗體,。
更有意義的是,，檢測報告基因的表達情況可以用來實時追蹤流感病毒在動物模型體內的感染、生長與分布,。因此，該成果不但為流感病毒的基礎研究,，也為流感疫苗,、抗流感病毒藥物和抗體的研發(fā)提供了強有力的工具,。
此項研究得到了中國科學院、國家自然科學基金,、呼吸疾病國家重點實驗室,、廣東省自然科學基金等資助。（生物谷Bioon.com）
生物谷推薦原文出處：
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.
