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草地贪夜蛾Spdoptera frugiperda是危害玉米、高粱等农作物的重要害虫。草地贪夜蛾卵巢细胞系IPLB-Sf-9(Sf9)广泛应用于病毒学研究和真核生物基因表达。本研究从Sf9细胞中提取总RNA,应用SMARTTM技术,构建以pGADT7-Rec为载体的Sf9细胞酵母GAL4AD融合cDNA文库。Sf9细胞总RNA的电泳谱型与哺乳动物总RNA存在差异,其28S rRNA带弱于18S rRNA带,所合成的原始ds cDNA大小为0.1～4kb。文库展现良好的多态性,载体质粒插入的cDNA片段大小大部分在0.3～2kb之间。Sf9细胞酵母GAL4AD融合cDNA文库的构建为草地贪夜蛾功能基因的识别和鉴定,为研究杆状病毒与宿主细胞的互作机制提供了重要研究工具。
Spodoptera frugiperda Spodoptera frugiperda is an important pest damage to corn, sorghum and other crops. Spodoptera frugiperda ovarian cell line IPLB-Sf-9 (Sf9) is widely used in virological research and eukaryotic gene expression. In this study, total RNA was extracted from Sf9 cells and SMARTTM technology was used to construct a yeast GAL4AD fusion cDNA library of Sf9 cells transfected with pGADT7-Rec. The total RNA of Sf9 cells was different from the total RNA of mammals. The 28S rRNA band of Sf9 cells was weaker than that of 18S rRNA. The size of original ds cDNA was 0.1-4 kb. The library showed good polymorphism, most of the inserted cDNA fragments were between 0.3 ~ 2kb in size. Construction of Sf9 Cell Yeast GAL4AD Fusion cDNA Library The identification and characterization of functional genes of S. frugiperda provided an important research tool for studying the interaction mechanism between baculovirus and host cells.