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Dynamitin真核表达载体的构建及鉴定 被引量：1

Construction of eukaryotic expression plasmid expressing dynamitin gene
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摘要 目的构建pRe-dyn真核表达质粒,为研究dynein-dynactin复合体在登革病毒逆行性运输中的作用奠定基础。方法RT-PCR扩增dynamitin基因片段后,将其克隆入真核表达载体pReceiver-M01a;通过PCR、酶切、测序和间接免疫荧光染色鉴定重组质粒。结果经PCR扩增、酶切和测序验证,重组质粒构建正确,命名为pRe-dyn;重组质粒转染Vero细胞后,间接免疫荧光染色显示胞浆中有dyanimtin蛋白的表达。结论成功构建了真核表达质粒pRe-dyn,为后续研究dynamitin在病毒感染过程中的作用积累了有价值的实验资料。 Objective To construct a recombinant plasmid expressing dynamitin gene for studying the role of dynein-dynactin in the retrograde transport of Dengue virus. Methods The genc fragment of dynamifin was obtained by RT-PCR and cloned into an eukaryotic expression vector pReceiver-M01a. Then the recombinant plasmid was confirmed by PCR, restriction endonuclease digestion, sequencing, and indirect immunofluorescence assay. Results The recombinant plasmid, named pRe-dyn, was constructed and identified. Meanwhile, Vero cells transfected with the plasmid showed reactivity with antibody against dyanimntin in the cytoplasmic region. Conclusion The plasmid pRe-dyn has been constructed succesfully, which lays a base for further study on roles of dynamitin in virus infection.
作者 陈炜 高娜 王嘉丽 田衍平 陈宗涛 徐小峰 张俊磊 刘丽梅 江雯 安静
机构地区 第三军医大学基础医学部微生物学教研室重庆市微生物工程实验室 第三军医大学基础医学部组织胚胎学教研室 首都医科大学基础医学院微生物学教研室
出处 《免疫学杂志》 CAS CSCD 北大核心 2008年第2期167-169,共3页 Immunological Journal
基金 国家自然科学基金(30471552,30640065和30671853)
关键词 病毒 微管骨架 逆行性运输 胞质动力蛋白 Dynamitin Virus Microtubules Retrograde transport Cytoplasmic dynein Dynamitin
分类号 R346 [医药卫生—基础医学]
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参考文献7

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共引文献1

同被引文献8

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免疫学杂志
2008年 第2期

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