Showing 2 results for Transient Expression
Neda Mirakhorli, Afsane Heydari Reyhani, Behrooz Shiran, Sadollah Hooshmand,
Volume 3, Issue 1 (4-2014)
Abstract
since permanent transfer of genes and transgenic plant production, particularly in perennial plants such as almonds are time and cost consuming, optimization of various factors affecting the transgenic plants is critical. Transient genetic expression using Agrobacterium (Agroinfiltration) in plant is a useful tool to study gene function indifferent gene expression systems. Since optimization of factors that could affect the transgenic methods is necessary for gene transfer programs, Because This study was aimed at developing an efficient system for transient expression via Agrobacterium in almond leaves for rapid prediction of systems performance. Here the lichenasereporter gene was usedto assessthe parametersinfluencing thelevel oftransient expression. The main advantagesof using thisgene includeeasy operation, high sensitivity and biosafety. Three different constructs carrying lichenase gene and differentregulatoryelementswereinfiltrated intoleaves ofmamaei and shahrood 12cultivars,by two Agrobacteriumstrain, LBA4404 andGV3501,using syringewithoutneedle.After threedays,the amount of active proteinwas measured. Statistical analysis showedthat the highestamount ofprotein was obtainedin theleavesof mamaei cultivar transformedby theexpression systemcausing accumulation ofproteins in theendoplasmic reticulum by LBA4404 strain. So forrapidproduction ofrecombinant protein in Almond, theagroinfiltration method by using the related expression system, LBA4404 strain and mamaei cultivar, isrecommended.
Sahba Toosi, Farhad Shokouhifar, Saeid Malekzadeh Shafaroudi, Abdolreza Bagheri,
Volume 3, Issue 2 (10-2014)
Abstract
transient expression of a candidate effector gene in host plants following Agroinfiltration is a useful method to investigate the role of the gene in pathogenecity. In this study, the effects of two Agrobacterium tumefaciens strains on four standard melon lines and two Iranian melon cultivars were investigated to establish transient expression in melon. Compatibility of the LBA4404 and GV3101 strains was defined by injection of the bacterial suspensions into the leaves of cultivars. Viability of the injected Agrobacterium strain cells in leaf tissues was evaluated 24 hours after injection by prokaryotic GUS reporter gene assay. The LBA4404 strain harboring the pCAMBIA3301 vector containing an intron-GUS reporter gene was used to confirm eukaryotic GUS expression in the plant cells. The LBA4404 strain was transformed by the pBI121 and pCAMBIA3301 expression vectors and transient transformation was confirmed by colony PCR technique using the PSh3-F/R primers. The efficiency of transient transformation of the melon leaves was evaluated 48 hours after Agroinjection of the LBA4404 strain containing pCAMBIA3301 by the histochemical GUS assay. The leaves that were injected by LBA4404 harboring pBI121 showed GUS activity. All the melon cultivars transiently expressed the GUS reporter gene 24 hours after Agroinjection. These findings could be used in the future studies to evaluate function of candidate effector genes in interaction with standard melon lines.
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