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昆虫细胞的转录组分析以促进杆状病毒-昆虫表达。

Transcriptome analyses of insect cells to facilitate baculovirus-insect expression.

作者信息

Yu Kai, Yu Yang, Tang Xiaoyan, Chen Huimin, Xiao Junyu, Su Xiao-Dong

机构信息

Biodynamic Optical Imaging Center, School of Life Science, Peking University, Beijing, 100871, China.

State Key Laboratory of Protein and Plant Gene Research, Peking University, Beijing, 100871, China.

出版信息

Protein Cell. 2016 May;7(5):373-82. doi: 10.1007/s13238-016-0260-y. Epub 2016 Mar 26.

Abstract

The High Five cell line (BTI-TN-5B1-4) isolated from the cabbage looper, Trichoplusia ni is an insect cell line widely used for baculovirus-mediated recombinant protein expression. Despite its widespread application in industry and academic laboratories, the genomic background of this cell line remains unclear. Here we sequenced the transcriptome of High Five cells and assembled 25,234 transcripts. Codon usage analysis showed that High Five cells have a robust codon usage capacity and therefore suit for expressing proteins of both eukaryotic- and prokaryotic-origin. Genes involved in glycosylation were profiled in our study, providing guidance for engineering glycosylated proteins in the insect cells. We also predicted signal peptides for transcripts with high expression abundance in both High Five and Sf21 cell lines, and these results have important implications for optimizing the expression level of some secretory and membrane proteins.

摘要

从甘蓝夜蛾(Trichoplusia ni)中分离得到的High Five细胞系(BTI-TN-5B1-4)是一种广泛用于杆状病毒介导的重组蛋白表达的昆虫细胞系。尽管它在工业和学术实验室中得到了广泛应用,但其基因组背景仍不清楚。在此,我们对High Five细胞的转录组进行了测序,并组装了25,234个转录本。密码子使用分析表明,High Five细胞具有强大的密码子使用能力,因此适合表达真核和原核来源的蛋白质。我们的研究对参与糖基化的基因进行了分析,为在昆虫细胞中工程化糖基化蛋白提供了指导。我们还预测了在High Five和Sf21细胞系中高表达丰度转录本的信号肽,这些结果对于优化某些分泌蛋白和膜蛋白的表达水平具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6e6e/4853316/188ed6a18b39/13238_2016_260_Fig1_HTML.jpg

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