Transient gene expression and stable genetic transformation into conifer tissues by microprojectile bombardment

1996 ◽  
pp. 417-462 ◽  
Author(s):  
Armand Séguin ◽  
Denis Lachance ◽  
Pierre J. Charest
Keyword(s):  
Gene Expression ◽  
Genetic Transformation ◽  
Microprojectile Bombardment ◽  
Transient Gene Expression

Transient gene expression in sunflower (Helianthus annuus L.) following microprojectile bombardment

Plant Science ◽  
1995 ◽  
Vol 105 (1) ◽  
pp. 95-109 ◽  
Author(s):  
Reiner Hunold ◽  
Monique Burrus ◽  
Roberte Bronner ◽  
Jean-Pierre Duret ◽  
Günther Hahne
Keyword(s):  
Gene Expression ◽  
Helianthus Annuus ◽  
Microprojectile Bombardment ◽  
Transient Gene Expression ◽  
Helianthus Annuus L

scholarly journals Production of transgenic bean callus via genetic transformation by DNA-coated tungsten particles

2003 ◽  
Vol 9 (3-4) ◽  
Author(s):  
E. Eissa Ahmed
Keyword(s):  
Gene Expression ◽  
Genetic Transformation ◽  
Transient Gene Expression ◽  
Callus Cultures ◽  
Ms Medium ◽  
Mannitol Dehydrogenase ◽  
Dehydrogenase Gene ◽  
Half Strength ◽  
Media Data ◽  
Callus Lines

Callus cultures were induced from hypocotyl of young bean seedlings. The B5 medium completed with 1 mg/1 KIN and 2mg/1 2,4-D proved the best. Callus developed and maintenaned on B5 medium supplemented with 1mg/1 kinetin and 2mg/I 2,4-D. The B5 medium supplemented with 1mg/1 KIN and 2mg/1 2,4-D induced much more callus than half strength MS medium supplemented with 0.5 or 0.75mg/1 BA and 0.1 mg/1 NAA. The results demonstrate that GeneboosterTM is convenient method to obtain transient gene expression in callus of bean. The results have shown that the bean callus shot by GeneboosterTM can be transformed to get (kanamycin-resistant and stress mannitol­tolerant) calli. The presence of mannitol-dehydrogenase gene (mt/) was verified by PCR, showing the integration of mt/ gene carried by two plasmids. Co-transformed calli were selected after bombardment on kanamycin, mannitol and (kanamycin+mannitop-containing media. Data of molecular analysis (PCR) confirmed the insertion of mtl gene in the genome of mannitol-tolerant callus lines.

scholarly journals Identification of Citrus Fruit-Specific and Pathogen-Induced Promoters and Their Use in Molecular Engineering

2001 ◽  
Author(s):  
Ron Porat ◽  
Doron Holland ◽  
Linda Walling
Keyword(s):  
Gene Expression ◽  
Promoter Activity ◽  
Shoot Growth ◽  
Microprojectile Bombardment ◽  
Transient Gene Expression ◽  
Citrus Fruit ◽  
Gus Gene ◽  
Promoter Fragment ◽  
The Us ◽  
Gus Gene Expression

This one year BARD project was funded to develop methods to monitor promoter activity a gene expression patterns in citrus fruit. To fulfill this goal, we divided the research tasks between both labs so that the Israeli side evaluated the use of microprojectile bombardment ; a tool to evaluate transient gene expression in various citrus fruit tissues, and the US side optimized technical parameters required for Agrobacterium-mediated transformation of various citrus cultivars. Microprojectile bombardment appeared to be a very efficient method for transient gene expression analysis in citrus leaf tissues but was somewhat less applicable in fruit tissues. Nevertheless, we did succeeded to achieve significant levels of 35S-GUS gene expression in young green flavedo tissue. However, only single random spots of 35S-GUS gene expression were detected mature flavedo and in juice sacs and albedo tissue. Overall, we assume that following some more technical improvements particle bombardment could provide a useful technique to rapidly analyze promoter activity at least in the flavedo tissue. For Agrobacterium-mediated transformation, we found that shoot cultures of 'Washington' navel oranges,'Fairchild' mandarins,'Eureca' lemons,'Troyer' citrange and various grapefruits provided a more reliable and consistent source of tissue for transformation than germinated seedlings. Moreover, various growth media's (McCown, Quoirin & Lepoivre, DCR) further improved shoot and root growth relative to MS mineral media, which is commonly used. Also pure white light (using bulbs which do not emit UV or blue light) improved shoot growth in various citrus varieties, and paromomycin appeared to be a more efficient antibiotic for the selection of transgenic plants than Kanamycin. Overall, these optimizations improve transformation efficacy and shoot growth and rooting capacity. In addition to the development of transformation methods, both Israeli and US labs achieved progress in the identification of citrus fruit-specific promoters. In Israel, we isolated a 3.6 kb promoter fragment of the thiamine biosynthesis c-thi gene, which is highly expressed in fruit peel tissue, whereas in the US we isolated a 1.5 kb promoter fragment of the citrus seed-specific cDNA CssH. The identification of more fruit-specific cDNAs and their corresponding promoter regions is currently in progress.

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.

Poly(ethyleneimine)-Mediated Large-Scale Transient Gene Expression: Influence of Molecular Weight, Polydispersity and N -Propionyl Groups

2012 ◽  
Vol 12 (5) ◽  
pp. 628-636 ◽  
Author(s):  
Zuzana Kadlecova ◽  
Sophie Nallet ◽  
David L. Hacker ◽  
Lucia Baldi ◽  
Harm-Anton Klok ◽  
...  
Keyword(s):  
Gene Expression ◽  
Molecular Weight ◽  
Large Scale ◽  
Transient Gene Expression

scholarly journals Mild Hypothermia Improves Transient Gene Expression Yields Several Fold in Chinese Hamster Ovary Cells

2008 ◽  
Vol 24 (2) ◽  
pp. 458-465 ◽  
Author(s):  
S. Wulhfard ◽  
S. Tissot ◽  
S. Bouchet ◽  
J. Cevey ◽  
M. DeJesus ◽  
...  
Keyword(s):  
Gene Expression ◽  
Chinese Hamster Ovary ◽  
Mild Hypothermia ◽  
Chinese Hamster Ovary Cells ◽  
Transient Gene Expression ◽  
Chinese Hamster ◽  
Ovary Cells
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