| HIV-1B亚型假病毒的制备与鉴定 |
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| 引用本文： | 王茂鹏,李昌,杜寿文,朱羿龙,朱娜,孙丹丹,金宁一. HIV-1B亚型假病毒的制备与鉴定[J]. 军事医学科学院院刊, 2014, 0(1): 31-34 |
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| 作者姓名： | 王茂鹏 李昌 杜寿文 朱羿龙 朱娜 孙丹丹 金宁一 |
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| 作者单位： | [1]吉林大学动物医学学院,长春130062 [2]军事医学科学院军事兽医研究所,省部共建吉林省人兽共患病预防与控制重点实验室,长春130122 |
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| 基金项目： | [基金项目]国家973计划前期研究资助项目（2011CB512110）;国家自然科学基金资助项目（81001342）;中国博士后科学基金面上资助项目（20110491803） |
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| 摘 要： | 目的：制备携带增强绿色荧光蛋白（ EGFP）基因和ENV包膜蛋白的HIV-1 B亚型假病毒用于感染研究，并建立可行的鉴定方法。方法双质粒共转染HEK293 T细胞后收获病毒上清， TRIzol法提取病毒基因组并采用RT-PCR进行报告基因扩增，Western印迹和ELISA法检测病毒P24抗原。假病毒感染HIV-1允许细胞，进行报告基因检测、病毒滴度测定以及单轮感染活性实验研究。结果与结论建立了HIV-1假病毒制备与鉴定方法，制备获得了B型HIV-1假病毒，经鉴定具备感染SupT1和TZM-bl细胞能力，为HIV-1与宿主细胞相互作用研究奠定了基础。
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| 关 键 词： | HIV-1 B亚型假病毒 感染 鉴定方法 |
Preparation and identification of Env-pseudotyped subtype B HIV-1 |
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| WANG Mao-peng,LI Chang,DU Shou-wen,ZHU Yi-long,ZHU Na,SUN Dan-dan,JIN Ning-yi. Preparation and identification of Env-pseudotyped subtype B HIV-1[J]. Bulletin of the Academy of Military Medical Sciences, 2014, 0(1): 31-34 |
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| Authors: | WANG Mao-peng LI Chang DU Shou-wen ZHU Yi-long ZHU Na SUN Dan-dan JIN Ning-yi |
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| Affiliation: | 1. College of Veterinary Medieine, Jilin University, Changehun 130062, China;2. Key Laboratory of Zoonosis Prevention and Control of Jilin Provinee, Institute of Military Veterinary, Academy of Military Medieal Seienees, Changehun 130122, China) |
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| Abstract: | Objective To prepare the Env-pseudotyped subtype B HIV-1 with enhanced green fluorescent protein （ EG-FP） gene,explore HIV-1 infection mechanisms and develop feasible methods of identification .Methods The Env-pseudo-typed viruses were packaged in HEK293T cells by cotransfection, and the reporter gene and P24 protein were detected by PCR, Western blot and ELISA .Reporter gene amplification , viral titration assay and a single round of infection assay were performed after the env-pseudotyped viruses infected HIV-1 permissive cell .Results and Conclusion A generation and identification method of the pseudotyped HIV-1 was established . The Env-pseudotyped subtype B HIV-1 has been prepared, which is able to infect SupT1 and TZM-bl cells through infection assay . |
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| Keywords: | Env-pseudotyped subtype B HIV-1 infection identification |
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