犬贾第虫病毒转染载体介导绿色荧光蛋白在该寄生虫中的表达
[Giardia canis virus transfection vector-mediated expression of green fluorescent protein in the parasite].
作者信息
Chen Li-Feng, Li Jian-Hua, Liu Quan, Zhao Yue-Ping, Cao Li-Li, Zhang Xi-Chen
机构信息
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China.
出版信息
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2007 Feb 28;25(1):36-40.
OBJECTIVE
To construct Giardia canis virus (GCV) transfection vector.
METHODS
According to transcriptional start site, replication origin and packaging site of GCV genome (DQ238861), a system was developed for the expression of a foreign gene in this organism by flanking the green fluorescent protein (GFP) gene with the fragments of GCV positive-strand RNA. The transcript of the construct was synthesized in vitro with T7 RNA polymerase and used to transfect GCV-infected trophozoites by electroporation.
RESULTS
The recombinant plasmid pGCV634/GFP/GCV2174 was constructed. The expression of green fluorescent protein mediated by GCV transfection vector in Giardia canis peaked at 1 d after electroporation (A490=1.8), and slowly decreased until 14 d post-transfection.
CONCLUSION
The engineered GCV vector can be successfully used to introduce and efficiently express a heterologous gene in the eukaryotic microorganism.
目的
构建犬贾第虫病毒（GCV）转染载体。
方法
根据GCV基因组（DQ238861）的转录起始位点、复制起点和包装位点，开发了一种通过用GCV正链RNA片段侧翼连接绿色荧光蛋白（GFP）基因，在该生物体中表达外源基因的系统。用T7 RNA聚合酶体外合成构建体的转录本，并通过电穿孔用于转染GCV感染的滋养体。
结果
构建了重组质粒pGCV634/GFP/GCV2174。GCV转染载体介导的绿色荧光蛋白在犬贾第虫中的表达在电穿孔后1天达到峰值（A490 = 1.8），并在转染后14天缓慢下降。
结论
工程化的GCV载体可成功用于在真核微生物中导入并高效表达异源基因。
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