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细胞内定位和基因表达分析为脱水生物细胞系中LEA蛋白的多样性提供了新见解。

Intracellular Localization and Gene Expression Analysis Provides New Insights on LEA Proteins' Diversity in Anhydrobiotic Cell Line.

作者信息

Kondratyeva Sabina A, Voronina Taisiya A, Nesmelov Alexander A, Miyata Yugo, Tokumoto Shoko, Cornette Richard, Vorontsova Maria V, Kikawada Takahiro, Gusev Oleg A, Shagimardanova Elena I

机构信息

Regulatory Genomics Research Center, Institute of Fundamental Medicine and Biology, Kazan Federal University, 420012 Kazan, Russia.

Division of Biomaterial Science, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba 305-0851, Japan.

出版信息

Biology (Basel). 2022 Mar 22;11(4):487. doi: 10.3390/biology11040487.

Abstract

Anhydrobiosis, an adaptive ability to withstand complete desiccation, in the nonbiting midge , is associated with the emergence of new multimember gene families, including a group of 27 genes of late embryogenesis abundant (LEA) proteins (). To obtain new insights into the possible functional specialization of these genes, we investigated the expression and localization of genes in a -derived cell line (Pv11), capable of anhydrobiosis. We confirmed that all but two genes identified in the genome of are expressed in Pv11 cells. Moreover, genes are induced in Pv11 cells in response to anhydrobiosis-inducing trehalose treatment in a manner highly similar to the larvae of during the real induction of anhydrobiosis. Then, we expanded our previous data on PvLEA proteins localization in mammalian cells that were obtained using C-terminal fusions of PvLEA proteins and green fluorescent protein (GFP). We investigated PvLEA localization using N- and C-terminal fusions with GFP in Pv11 cells and the Sf9 insect cell line. We observed an inconsistency of PvLEA localization between different fusion types and different cell cultures, that needs to be taken into account when using PvLEA in the engineering of anhydrobiotic cell lines.

摘要

在非吸血蠓中,隐生现象是一种耐受完全脱水的适应能力,它与新的多成员基因家族的出现有关,包括一组27个晚期胚胎发生丰富(LEA)蛋白基因()。为了深入了解这些基因可能的功能特化,我们研究了在能够进行隐生的源自的细胞系(Pv11)中这些基因的表达和定位。我们证实,在基因组中鉴定出的除两个基因外的所有基因都在Pv11细胞中表达。此外,在Pv11细胞中,基因会因诱导隐生的海藻糖处理而被诱导,其方式与在真正的隐生诱导过程中的幼虫高度相似。然后,我们扩展了我们之前关于PvLEA蛋白在哺乳动物细胞中定位的数据,这些数据是使用PvLEA蛋白与绿色荧光蛋白(GFP)的C端融合获得的。我们在Pv11细胞和Sf9昆虫细胞系中使用与GFP的N端和C端融合来研究PvLEA的定位。我们观察到不同融合类型和不同细胞培养物之间PvLEA定位存在不一致,在将PvLEA用于隐生细胞系工程时需要考虑到这一点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0dc4/9031878/0545abe28467/biology-11-00487-g001.jpg

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