Intracellularlocalization of an endogenous cellulosesynthase of Micrasterias denticulata(Desmidiales, Chlorophyta) by means oftransientgenetic transformation
Intracellularlocalization of an endogenous cellulosesynthase of Micrasterias denticulata(Desmidiales, Chlorophyta) by means oftransientgenetic transformation
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通过瞬时遗传转化对 Micrasterias denticulata（Desmidiales，Chlorophyta）内源纤维素合酶进行细胞内定位
DOI：
10.1111/j.1529-8817.2010.00867.x
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
W
中科院分区：
文献类型：
--
作者：
Vannerum;K.;Abe;J.;Sekimoto;H.;Inze,D.;Vyverman;W
细齿微鼓藻Micrasterias denticulataBréb.为研究链霉菌细胞壁生物学和形态学提供了有用的材料。然而，没有工具来分析细胞生物学过程中，在这个物种是可用的。在本研究中，实现了在Closterium peracerosum-strigosum-littoralecomplex（CpCAB 1）启动子的chla/b结合蛋白基因控制下，M. denticulata和说明的内源性纤维素合酶（MdCesA 1）的细胞内定位。在微粒轰击后实现了1/5，000细胞的转化效率。游离的绿色荧光蛋白（GFP）信号在细胞核和细胞质中均被检测到。另一方面，MdCesA 1-GFP融合蛋白以颗粒形式出现在质膜上，这些颗粒集中在叶的凹陷处、叶的尖端处，并且在较小程度上集中在叶的两侧沿着。因此，反映了细胞的多极生长机制。此外，边缘的细胞质隔间，最有可能的dictyosomes，标记，按照已知的分泌途径的纤维素合成酶复合物。除了细胞内定位研究，该系统的过表达表型的效用进行了讨论。
The desmidMicrasterias denticulataBréb. is useful for the study of streptophyte cell wall biology and morphology. However, no tools to analyze cell biological processes in vivo in this species are available. In the present study, transient gene expression under the control of the chla/b–binding protein gene of theClosterium peracerosum–strigosum–littoralecomplex (CpCAB1) promotor was achieved forM. denticulataand illustrated by the intracellular localization of an endogenous cellulose synthase (MdCesA1). A transformation efficiency of 1/5,000 cells was achieved following microparticle bombardment. The free green fluorescent protein (GFP) signal was detected both in the nucleus and in the cytoplasm. The MdCesA1‐GFP fusion protein, on the other hand, occurred at the plasma membrane in particles concentrated at the lobe indentations, the lobe tips, and, to a lesser extent, along the lobe sides. Hence, the multipolar growth mechanism of the cell is reflected. In addition, the margins of cytoplasmic compartments, most likely dictyosomes, were labeled, in accordance with the known secretory pathway of cellulose synthase complexes. Besides intracellular localization studies, the utility of the system for overexpression phenotyping is discussed.