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问号钩端螺旋体毒力相关蛋白InvA转录和表达特征的研究 被引量:2
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作者 罗依惠 陈铭 +2 位作者 李立伟 钱景 严杰 《中华微生物学和免疫学杂志》 CAS CSCD 北大核心 2009年第1期24-28,共5页
目的确定我国不同基因种问号钩端螺旋体(简称钩体)参考标准株携带毒力相关基因invA的情况,了解问号钩体赖株感染细胞前后invA基因转录和表达水平的变化。方法采用PCR检测4个不同基因种问号钩体株及双曲钩体PatocI株invA基因。克隆问... 目的确定我国不同基因种问号钩端螺旋体(简称钩体)参考标准株携带毒力相关基因invA的情况,了解问号钩体赖株感染细胞前后invA基因转录和表达水平的变化。方法采用PCR检测4个不同基因种问号钩体株及双曲钩体PatocI株invA基因。克隆问号钩体全长irwA基因并测序,构建问号钩体黄疸出血群赖型赖株invA基因原核表达系统。Ni-NTA亲和层析法提纯目的重组蛋白rInvA后免疫家兔获得抗血清,免疫双扩散法检测其效价。建立问号钩体赖株感染人胚肾上皮细胞HEK293模型,采用荧光定量RT—PCR和Westernblot分别检测问号钩体赖株感染HEK293细胞前后invA基因的转录和表达水平的变化。结果4个不同基因种的问号钩体株均含有invA基因,双曲钩体PatocI株则否。4株不同基因种的问号钩体irwA基因核苷酸和氨基酸序列相似性分别为99.33%~100%和98.66%~100%。所构建的原核表达系统能有效地表达rInvA。rInvA兔抗血清免疫双扩效价为1:16。问号钩体赖株感染HEK293细胞30min以后可大量黏附于细胞表面。问号钩体赖株感染HEK293细胞30min时,invA基因mRNA水平明显上调,45min时达到峰值,然后逐渐下降。问号钩体赖株感染HEK293细胞后45min和60min时可检出InvA蛋白,感染前及感染90min以后检测结果均为阴性。结论invA基因是致病性问号钩体所特有的基因。invA基因具有宿主细胞接触式表达及瞬时表达的特点,与问号钩体侵人宿主细胞密切相关。 展开更多
关键词 问号钩端螺旋体 侵入/感染 invA基因 转录/表达
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Viral infection of tobacco plants improves performance of Bemisia tabaci but more so for an invasive than for an indigenous biotype of the whitefly 被引量:15
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作者 Jian LIU Meng LI +4 位作者 Jun-min LI Chang-jun HUANG Xue-ping ZHOU Fang-cheng XU Shu-sheng LIU 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2010年第1期30-40,共11页
The ecological effects of plant-virus-vector interactions on invasion of alien plant viral vectors have been rarely investigated. We examined the transmission of Tomato yellow leaf cur/China virus (TYLCCNV) by the i... The ecological effects of plant-virus-vector interactions on invasion of alien plant viral vectors have been rarely investigated. We examined the transmission of Tomato yellow leaf cur/China virus (TYLCCNV) by the invasive Q biotype and the indigenous ZHJ2 biotype of the whitefly Bemisia tabaci, a plant viral vector, as well as the influence of TYLCCNV-infection of plants on the performance of the two whitefly biotypes. Both whitefly biotypes were able to acquire viruses from infected plants and retained them in their bodies, but were unable to transmit them to either tobacco or tomato plants. However, when the Q biotype fed on tobacco plants infected with TYLCCNV, its fecundity and longevity were increased by 7- and 1-fold, respectively, compared to those of the Q biotype fed on uninfected tobacco plants. When the ZHJ2 biotype fed on virus-infected plants, its fecundity and longevity were increased by only 2- and 0.5-fold, respectively. These data show that the Q biotype acquired higher beneficial effects from TYLCCNV-infection of tobacco plants than the ZHJ2 biotype. Thus, the Q biotype whitefly may have advantages in its invasion and displacement of the indigenous ZHJ2 biotype. 展开更多
关键词 Bemisia tabaci BIOTYPES BEGOMOVIRUS Vector-virus interaction Biological invasion
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