scholarly journals Organelle Visualization With Multicolored Fluorescent Markers in Bamboo

2021 ◽  
Vol 12 ◽  
Author(s):  
Mengdi Zhang ◽  
Shuai Hu ◽  
Fang Yi ◽  
Yanli Gao ◽  
Dongmei Zhu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Subcellular Localization ◽  
Molecular Mechanisms ◽  
Expression System ◽  
Transient Gene Expression ◽  
Moso Bamboo ◽  
Functional Study ◽  
Efficient System ◽  
Gene Expression System ◽  
Fluorescent Markers

Bamboo is an important model plant to study the molecular mechanisms of rapid shoot growth and flowering once in a lifetime. However, bamboo research about protein functional characterization is largely lagged behind, mainly due to the lack of gene transformation platforms. In this study, a protoplast transient gene expression system in moso bamboo has been first established. Using this reliable and efficient system, we have generated a set of multicolored fluorescent markers based on the targeting sequences from endogenous proteins, which have been validated by their comparative localization with Arabidopsis organelle markers, in a combination with pharmaceutical treatments. Moreover, we further demonstrated the power of this multicolor marker set for rapid, combinatorial analysis of the subcellular localization of uncharacterized proteins, which may play potential functions in moso bamboo flowering and fast growth of shoots. Finally, this protoplast transient gene expression system has been elucidated for functional analysis in protein–protein interaction by fluorescence resonance energy transfer (FRET) and co-immunoprecipitation analysis. Taken together, in combination with the set of moso bamboo organelle markers, the protoplast transient gene expression system could be used for subcellular localization and functional study of unknown proteins in bamboo and will definitely promote rapid progress in diverse areas of research in bamboo plants.

scholarly journals Harnessing a Transient Gene Expression System in Nicotiana benthamiana to Explore Plant Agrochemical Transporters

Plants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 524
Author(s):  
Bingqi Wu ◽  
Zhiting Chen ◽  
Xiaohui Xu ◽  
Ronghua Chen ◽  
Siwei Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transient Expression ◽  
Nicotiana Benthamiana ◽  
Heterologous Gene Expression ◽  
Expression System ◽  
Functional Characterization ◽  
Transient Gene Expression ◽  
High Mobility ◽  
Gene Expression System

Functional characterization of plant agrichemical transporters provided an opportunity to discover molecules that have a high mobility in plants and have the potential to increase the amount of pesticides reaching damage sites. Agrobacterium-mediated transient expression in tobacco is simple and fast, and its protein expression efficiency is high; this system is generally used to mediate heterologous gene expression. In this article, transient expression of tobacco nicotine uptake permease (NtNUP1) and rice polyamine uptake transporter 1 (OsPUT1) in Nicotiana benthamiana was performed to investigate whether this system is useful as a platform for studying the interactions between plant transporters and pesticides. The results showed that NtNUP1 increases nicotine uptake in N. benthamiana foliar discs and protoplasts, indicating that this transient gene expression system is feasible for studying gene function. Moreover, yeast expression of OsPUT1 apparently increases methomyl uptake. Overall, this method of constructing a transient gene expression system is useful for improving the efficiency of analyzing the functions of plant heterologous transporter-encoding genes and revealed that this system can be further used to study the functions of transporters and pesticides, especially their interactions.

scholarly journals Transient Gene Expression System Using Protoplasts from Developing Soybean Cotyledons.

1993 ◽  
Vol 43 (1) ◽  
pp. 53-60
Author(s):  
Mayuko ISHIKAWA ◽  
Kyuya HARADA ◽  
Fukumi SAKAI ◽  
Yuko OHASHI ◽  
Tadao NAITO
Keyword(s):  
Gene Expression ◽  
Expression System ◽  
Transient Gene Expression ◽  
Gene Expression System

An Agrobacterium-mediated transient gene expression system for intact leaves

Plant Science ◽  
1997 ◽  
Vol 122 (1) ◽  
pp. 101-108 ◽  
Author(s):  
Jyoti Kapila ◽  
Riet De Rycke ◽  
Marc Van Montagu ◽  
Geert Angenon
Keyword(s):  
Gene Expression ◽  
Expression System ◽  
Transient Gene Expression ◽  
Gene Expression System ◽  
Intact Leaves

scholarly journals Enhancement of transient erythropoietin protein expression by valproic acid in CHO‐K1 suspension adapted cells

2020 ◽  
Vol 25 (1) ◽  
pp. 28
Author(s):  
Yana Rubiyana ◽  
Retno Damajanti Soejoedono ◽  
Adi Santoso
Keyword(s):  
Gene Expression ◽  
Valproic Acid ◽  
Protein Expression ◽  
Histone Deacetylase Inhibitors ◽  
Expression System ◽  
Transient Gene Expression ◽  
Optimum Concentration ◽  
Enzyme Linked Immunosorbent Assay ◽  
Stable Gene ◽  
Gene Expression System

Erythropoietin (EPO) is a therapeutic protein that is widely used to increase red blood cell production in chronic kidney failure. EPO protein can be produced quickly with a transient gene expression system (TGE). However, the titer produced using TGE is usually lower than the stable gene expression system (SGE). It has been known that TGE can be improved by histone deacetylase inhibitors (iHDACs) such as valproic acid (VPA). This study was conducted to examine the VPA effect on EPO protein expression in CHO‐K1 suspension adapted cells and to find the optimum concentration of VPA on transient EPO protein production. EPO proteins was quantified using the enzyme‐linked immunosorbent assay (ELISA) method. The optimization of VPA concentrations showed that VPA increased the EPO protein yield by up to 2‐fold in transient EPO production, and the optimum concentration of VPA was 4 mM. VPA optimization was very helpful to obtain the maximum increase in the transiently expressed protein. Furthermore, this study can be used as a model to produce EPO proteins or other recombinant proteins rapidly with TGE of CHO‐K1 suspension adapted cells.

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