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[通过与绿色荧光蛋白报告基因的翻译融合确定拟南芥中PELPK1的亚细胞定位]

[Sub-celluiar localization of PELPK1 in Arabidopsis thaliana as determined by translational fusion with green fluorescent protein reporter].

作者信息

Rashid A

出版信息

Mol Biol (Mosk). 2014 Mar-Apr;48(2):300-5.

Abstract

PELPK1, a novel Arabidopsis thaliana gene was earlier annotated to encode a protein of sub-family, PELPK under hydroxyproline-rich glycoprotein (HRGP) super-family of proteins. Previous bioinformatics and computational analyses predicted PELPK1 to contain an amino-terminal signal peptide destined towards the secretory pathway. In the present study, transgenic plants were developed harboring a translational fusion construct comprising of PELPK1 coding sequence (PELPK1-CDS) and green fluorescent protein (GFP) reporter to determine the localization of PELPK1 in Arabidopsis plants. By employing the techniques of confocal laser scanning microscopy, immunolabeling of GFP with quantum dot (Q-dot), and transmission electron microscopy (TEM), it is shown that the translational fusion product is predominantly deposited to the cell wall. These results are in agreement with the earlier bioinformatics prediction that the PELPK1 is transported via the secretory pathway.

摘要

PELPK1是一种新的拟南芥基因,早期注释为在富含羟脯氨酸糖蛋白(HRGP)蛋白质超家族下编码PELPK亚家族的一种蛋白质。先前的生物信息学和计算分析预测PELPK1含有一个指向分泌途径的氨基末端信号肽。在本研究中,构建了一个包含PELPK1编码序列(PELPK1-CDS)和绿色荧光蛋白(GFP)报告基因的翻译融合载体,并培育出转基因植物,以确定PELPK1在拟南芥植物中的定位。通过共聚焦激光扫描显微镜技术、用量子点(Q点)对GFP进行免疫标记以及透射电子显微镜(TEM)观察,结果表明翻译融合产物主要沉积在细胞壁上。这些结果与早期生物信息学预测一致,即PELPK1通过分泌途径运输。

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