南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (03): 342-.

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从白纹伊蚊与致倦库蚊分离和鉴定可结合登革Ⅱ病毒的蛋白分子

付宇姣,郑学礼,潘京   

  • 出版日期:2013-03-20 发布日期:2013-03-20

Identification of dengue II virus-binding proteins from Aedes albopictus and Culex. Quinguefasciatus

  • Online:2013-03-20 Published:2013-03-20

摘要: 目的筛选登革Ⅱ病毒(DENV-2)识别白纹伊蚊、致倦库蚊的连接分子。方法提取不同发育时期白纹伊蚊、致倦库蚊的总
蛋白,用SDS-12% PAGE进行分析,用病毒覆盖蛋白结合试验(VOPBA)筛选不同发育时期白纹伊蚊、致倦库蚊表达连接
DENV-2分子。结果VOPBA检测不同发育时期白纹伊蚊显示:幼虫样品的泳道可见到25 000、35 000、50 000的DEN-2结合分
子,蛹样品的泳道可见到35 000、50 000分子,雄蚊样品可见到50 000分子,雌蚊样品中可见到35 000、50 000分子。DEN-2结
合蚊35 000分子在白纹伊蚊幼虫、蛹及成蚊的雌蚊的三个阶段均存在,而雄蚊的成蚊则缺少这一分子。VOPBA检测不同发育
时期致倦库蚊:幼虫样品可见100 000分子,蛹样品可见40、100 000分子,另外在50 000位置上下亦可见48 000、60 000左右的
多个条带,雄蚊与雌蚊样品均见40 000、100 000分子。结论35 000大小的蛋白分子分布与白纹伊蚊感染组织向性相关。

Abstract: Objective To screen DENV-2 binding proteins from Aedes albopictus and Culex. quinquefasciatus. Methods The total
proteins of Aedes albopictus and Culex. quinquefasciatus in different developmental stages were prepared and analyzed with
SDS-12% polyacrylamide gel. After electophoresis the proteins were transferred using Mini Trans-Blot Electrophoretic
Transfer Cell (Bio-Rad ) to a nitrocellulose membrane. Virus overlay protein-binding assay (VOPBA) was carried out using
anti-dengue virus 1-4 monoclonal antibody. Results In Aedes albopictus, VOPBA detected DEN-2 binding molecules of 25 000,
35 000, and 50 000 in larvae samples, molecules of 35 000 and 50 000 in pupae samples, a 50 000 molecule in male mosquito
samples, and molecules of 35 000 and 50 000 in female mosquito samples. DENV-2 binding protein of 35 000 was found in the
larvae, pupae, and female mosquitoes, but not in male mosquitoes. In Culex. Quinquefasciatus, VOPBA detected a molecule of
100 000 in larvae samples, molecules of 40 000, 100 000, and around 50 000 (48 000 and 60 000) in pupae samples, and
molecules of 40 000 and 100 000 in male mosquitoes and female mosquito samples. Conclusion Several proteins capable of
binding DENV are found in Aedes albopictus and Culex. quinquefasciatus in different development stages. The 35 000 molecule
expressed in Aedes albopictus as a putative receptor protein may be related to virus tropism in mosquito tissues.