Current Opinion in Neurobiology. 231-239 - The neurobiology

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S. Nelson, unpublished data). 1. Inoculation Methods Inoculum for the older BMV vectors developed by Ding et al. (21, 33) and Pacak et al. (22) is produced by in vitro transcription of linearized plasmids containing the BMV genome sequences. The transcripts are capped during the reaction, and the product is mixed with FES buffer and then rub-inoculated to seedlings (21). We determined that inoculation of the BMV vector to N. benthamiana first, as an intermediate host, provides a high titer of virus in extract from these plants for subsequent inoculation to the grass host (33).

Benthamiana intermediate host that were destined for the grass host. 3. RTBV-VIGS Vector RTBV is a double-stranded DNA virus of the genus Pararetrovirus. It has a genome of approximately 8 kb encoding four ORFs (I to IV). RTBV was modified to serve as a VIGS vector by assembling the viral DNA molecule as a partial dimer within the T-DNA of a binary plasmid. In addition, the RTBV promoter was replaced with the constitutively expressed maize ubiquitin promoter, and a tRNA-binding site and a multiple-cloning site, the latter for the insertion of foreign gene fragments, were added.

Zhang L, Lavery L, Gill U et al (2009) A cation/proton-exchanging protein is a candidate for the barley NecS1 gene controlling necrosis and enhanced defense response to stem rust. Theor Appl Genet 118:385–397 38. Hein I, Pacak MB, Hrubikova K et al (2005) Virus-induced gene silencing-based functional characterization of genes associated with powdery mildew resistance in barley. Plant Physiol 138:2155–2164 39. Shen QH, Saijo Y, Mauch S et al (2007) Nuclear activity of MLA immune receptors links isolate-specific and basal disease-resistance responses.

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