| 登革病毒3型病毒样颗粒在酵母体内的表达与鉴定 |
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| 引用本文： | 付春云,方丹云,刘岩,喻治准,江汉宁,江丽芳,周俊梅.登革病毒3型病毒样颗粒在酵母体内的表达与鉴定[J].热带医学杂志,2011,11(6):613-616,656. |
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| 作者姓名： | 付春云 方丹云 刘岩 喻治准 江汉宁 江丽芳 周俊梅 |
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| 作者单位： | 中山大学中山医学院医学微生物教研室,广东广州510080;中山大学热带病防治研究教育部重点实验室,广东广州,510080 |
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| 基金项目： | 国家自然科学基金(U0632002) |
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| 摘 要： | 目的 探讨登革病毒3型病毒样颗粒(DENV3-vLPs)的免疫原性.方法 用PCR法扩增登革病毒3型prME 基因,插入载体pGAPZaA构建重组质粒pGAPZaA-prME-D3,将其转化毕赤酵母X33构建转化子pGAPZaA-prME-D3/X33.对转化子表达上清和细胞裂解液进行SDS-PAGE和Western blotting分析.用蔗糖密度梯度离心纯化表达的DENV3-VLPs并进行鉴定和电镜观察.结果 成功构建重组载体pGAPZaA-prME-D3,获得酵母转化子pGAPZaA-prME-D3/X33,并应用毕赤酵母表达了DENV3-VLPs,表达蛋白约50 000 Mr,电镜观察vLPs直径为20～50 am.结论 应用酵母表达系统成功表达了DENV-3 VLPs,经鉴定表明其具有免疫反应性,为后续免疫原性研究及四价登革vLPs疫苗的构建奠定了基础.
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| 关 键 词： | 登革病毒 病毒样颗粒 蔗糖密度梯度离心 毕赤酵母 |
Expression and identification of dengue virus type 3 like particles in pichica pastoris |
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| FU Chun-yun,FANG Dan-yun,LIU Yan,YU Zhi-zhun,JIANG Han-ning,JIANG Li-fang,ZHOU Jun-mei.Expression and identification of dengue virus type 3 like particles in pichica pastoris[J].Journal Of Tropical Medicine,2011,11(6):613-616,656. |
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| Authors: | FU Chun-yun FANG Dan-yun LIU Yan YU Zhi-zhun JIANG Han-ning JIANG Li-fang ZHOU Jun-mei |
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| Institution: | FU Chun-yun,FANG Dan-yun,LIU Yan,YU Zhi-zhun,JIANG Han-ning,JIANG Li-fang,ZHOU Jun-mei(1.Department of Microbiology,Zhongshan School of Medicine,Sun Yat-sen University,Guangdong,Guangzhou 510080,2.Key Laboratory of Tropical Disease Control,Chinese Ministry of Education,China) |
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| Abstract: | Objective To construct DENV3-VLPs recombinant vector,purify and identify DENV3-VLPs expressed in P.pastoris expression system.Methods DEVN-3 prM and E genes were amplified by PCR and inserted into pGAPZaA to construct the recombinant plasmid pGAPZaA-prME-D3.The pGAPZaA-prME-D3 /X33 was constructed by transferring the plasmid to P.pastoris X33.The expression supernatant and cell lysate from pGAPZaA-prME-D3 /X33 was analyzed by SDS-PAGE and Western blotting.The sucrose density gradient ultracentrifugation was... |
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| Keywords: | dengue virus virus-like particles sucrose density gradient ultracentrifugation P pastoris |
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