Shang Yitong, Jin Qi, Huang Hui, Qin Kunhai, Yan Huanhuan, Hu Zhihong
Jiangxi Key Laboratory of Bioprocess Engineering, College of Life Science, Jiangxi Science & Technology Normal University, Nanchang 330013, Jiangxi, China.
Sheng Wu Gong Cheng Xue Bao. 2024 Sep 25;40(9):2998-3010. doi: 10.13345/j.cjb.240016.
The organelles in the multi-nucleated filamentous fungus present polymorphism. To observe the organelle morphology in . and provide references for the localization prediction of unknown proteins and the disclosure of biological reaction pathways in . , we fused different subcellular localization signals with green fluorescent protein (GFP) to obtain different subcellular localization vectors, which were then transferred into . by -mediated transformation. The . reporter strains with fluorescence-labeled nuclei, mitochondria, endoplasmic reticulum, vacuole, lipid droplets, peroxisome, and Golgi apparatus were successfully constructed. Furthermore, staining with small-molecule specific dyes was carried out to validate the co-localization of fluorescence-labeled mitochondria, nuclei, and lipid droplets in the reporter strains, which further confirmed that the reporter strains were successfully constructed. The distribution and morphology of fluorescence-labeled organelles were observed at different growth stages and under different culture conditions. The constructed reporter strains provide basic tools for studying the organelle morphology, localization of unknown target proteins, and subcellular localization in . .
多核丝状真菌中的细胞器呈现多态性。为了观察……中的细胞器形态，并为未知蛋白质的定位预测以及……中生物反应途径的揭示提供参考，我们将不同的亚细胞定位信号与绿色荧光蛋白（GFP）融合，以获得不同的亚细胞定位载体，然后通过……介导的转化将其转入……。成功构建了具有荧光标记的细胞核、线粒体、内质网、液泡、脂滴、过氧化物酶体和高尔基体的……报告菌株。此外，用小分子特异性染料进行染色，以验证报告菌株中荧光标记的线粒体、细胞核和脂滴的共定位，这进一步证实了报告菌株的成功构建。在不同生长阶段和不同培养条件下观察了荧光标记细胞器的分布和形态。构建的报告菌株为研究……中的细胞器形态、未知靶蛋白的定位和亚细胞定位提供了基础工具。
